LOCUS pHC-nCas9-Gam 7829 bp DNA circular SYN 14-SEP-2022 DEFINITION pHC-nCas9-Gam (F191): Ligation of GamApo-Spe-Bgl* into F190pHC-nCas9-UGI* ACCESSION pHC-nCas9-Gam KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct COMMENT DNA Cloning Service, https://dna-cloning.com/vectors/crispr/pHC-nCas9-Gam FEATURES Location/Qualifiers CDS 30..50 /label="O1" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" misc_feature 96..116 /label="NLS" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" misc_feature 117..134 /label="Linker" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 135..656 /label="Gam" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 657..704 /label="Linker" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 705..1388 /label="APOBEC1" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 1389..1484 /label="Linker" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 1485..5615 /label="Cas9D10A'" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 5616..5864 /label="UGI" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 5865..5894 /label="Linker" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 5895..6143 /label="UGI" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 6144..6155 /label="Linker" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 6156..6203 /label="NLS" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 6227..6247 /label="O2" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 6302..7162 /label="ORF-1" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 7165..7829 /label="ColE1" /note="color: #8B8F86" /ApEinfo_fwdcolor="#8B8F86" /ApEinfo_revcolor="#8B8F86" misc_feature 7387 /label="LC" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" ORIGIN 1 aacgccagca acgcggcctt tatacagata attgtgagcg gataacaatt aataggcgta 61 tcacgaggac tagtcgatcc tccataacta tggctcccaa gaagaagcgc aaagtcggca 121 tccacggagc tccaatggcc aagcctgcca agaggattaa gtccgcagcc gctgcctacg 181 tgccgcagaa ccgtgacgcg gttatcaccg acatcaagcg catcggtgat ctccaaagag 241 aggccagcag gctggagact gagatgaacg acgctattgc cgaaatcacc gagaagttcg 301 ccgctcgcat cgcacccatc aagaccgaca ttgagacact ctctaagggc gtccaggggt 361 ggtgcgaggc caatcgtgac gagcttacca acggcggaaa ggtcaagacc gccaacctcg 421 tgaccggcga tgtgtcctgg cgggttcgcc caccctcggt cagcatcagg ggcatggacg 481 ccgtgatgga aaccctggag cgcctcggat tgcagcgttt catcagaacc aagcaggaga 541 tcaacaagga ggctattctt ctggagccta aggccgtagc cggtgtcgct ggcatcactg 601 tgaaatcagg aatcgaggac ttctccatca ttccgtttga acaagaggcc gggatcagcg 661 gctccgagac tcccggaacc agtgagagcg cgactccgga atcctcttcc gagacaggcc 721 ccgtcgccgt ggacccaacc ctccgcaggc gaatcgagcc tcacgagttc gaagttttct 781 tcgatccccg cgagctgcgt aaggagactt gcctattgta cgagatcaac tggggtggca 841 ggcacagcat ttggcgccat acctcgcaga ataccaacaa gcacgtcgaa gtgaacttca 901 tcgagaagtt caccaccgag agatacttct gcccgaacac ccgttgctcc atcacttggt 961 tccttagctg gtcaccatgc ggagagtgct cccgcgctat caccgagttt ctgtctcgct 1021 acccacacgt caccctcttc atttacatcg ccaggctcta tcaccacgcc gaccctcgga 1081 accgtcaggg cctgcgcgac ttgatcagct ccggggtgac aatccaaatt atgaccgagc 1141 aggagtcggg atactgctgg aggaacttcg ttaattacag cccctccaac gaggcacact 1201 ggcccagata cccgcatctt tgggtccgcc tctacgtgct ggagctgtac tgcatcatct 1261 tgggccttcc accctgcctc aacatcctgc gtaggaagca gccccagctc accttcttca 1321 cgattgcctt gcaatcttgc cactaccagc gccttcctcc gcacatcctg tgggctaccg 1381 gtttgaagtc cggcggaagc agtggcgggt ccagcggctc agaaactcca ggaacctccg 1441 agagcgccac tcccgagtcg tccggcggtt ctagcggagg ctccgacaag aagtacagca 1501 ttggtctggc catcgggacc aactccgtcg gctgggccgt tatcaccgat gagtacaaag 1561 tgccttctaa gaagttcaaa gtcctcggaa acactgacag gcacagcatc aagaagaact 1621 tgattggcgc ccttctcttc gactccggcg aaaccgctga ggccacccgt ctgaagcgca 1681 ccgccagaag gcgctacacc cgtcgcaaga ataggatctg ctacctccag gagatcttca 1741 gcaacgagat ggccaaagtg gacgattcct tctttcaccg tttggaggag agcttccttg 1801 tcgaggagga caagaagcac gagcgccatc ccatcttcgg aaacattgtt gacgaggtgg 1861 cttaccacga gaagtatcca actatctacc acctcagaaa gaagctggtg gactccaccg 1921 acaaggccga tttgaggctc atctaccttg ccctcgctca catgatcaag ttccgcggtc 1981 acttcctgat tgagggcgac ctcaaccccg acaactctga cgtcgataag ttgttcatcc 2041 aactggtcca gacctacaac cagcttttcg aggagaatcc tatcaacgcc tccggggttg 2101 acgccaaggc tattctcagc gcccgtctgt ccaagagccg caggcttgag aacttgatcg 2161 cccagctccc aggagagaag aagaacggcc tcttcggaaa cctgatcgct ctctccttgg 2221 gccttacccc taacttcaag tctaattttg acctcgccga ggacgccaag ctgcaactca 2281 gcaaggacac ctacgatgac gacttggaca acctgctcgc ccagatcggc gatcagtacg 2341 ctgacctttt cttggccgcc aagaacctct ccgacgctat tctgctctcc gacatccttc 2401 gcgtgaacac tgagatcacc aaggccccac tgagcgcctc catgatcaag cgttacgacg 2461 aacaccacca agatttgacc ctcctcaagg ctcttgtcag acagcagctg cccgagaagt 2521 acaaggagat tttcttcgac cagagcaaga acggatacgc cggttacatc gacggcgggg 2581 cctcccaaga ggagttctac aagttcatca agcccatctt ggagaagatg gacggcaccg 2641 aagagctgct cgtgaagctc aaccgcgagg atcttttgag gaagcagcgt accttcgaca 2701 acggatctat tccccaccag atccatctcg gcgagctgca cgctatcctc cgcaggcaag 2761 aggacttcta tccattcctt aaggacaatc gcgagaagat tgaaaagatc ctcactttta 2821 gaatccctta ctacgtcgga cccttggccc gtggcaacag ccgcttcgcc tggatgacca 2881 ggaagtccga ggaaaccatc accccatgga acttcgagga ggtggttgac aagggggcca 2941 gcgctcagtc cttcattgag cgcatgacca acttcgataa gaacctgccc aacgagaaag 3001 tcctcccaaa gcactccctt ctgtacgagt acttcactgt ctacaatgag ttgaccaaag 3061 tgaagtacgt gaccgagggt atgcgtaagc ctgccttcct cagcggcgag cagaagaagg 3121 ccatcgttga cctcctgttc aagaccaaca ggaaagtcac cgtgaagcag cttaaggagg 3181 actacttcaa gaagatcgag tgctttgact ctgtcgagat cagcggagtg gaggatcgct 3241 tcaacgcttc cttgggcact taccacgacc tcctgaagat tatcaaggac aaggacttcc 3301 tcgacaacga ggagaacgaa gatatcttgg aggacatcgt tcttaccctc accctgttcg 3361 aggacagaga gatgattgag gaacgcctca agacctacgc ccacttgttc gacgataaag 3421 tcatgaagca acttaagcgt aggcgctaca ccggatgggg caggctgtcc cgtaagctca 3481 tcaacggtat ccgcgacaag cagagcggca agactattct cgacttcttg aagtccgacg 3541 gattcgccaa tcgcaacttc atgcagctga tccacgacga ttctcttacc ttcaaggagg 3601 acatccaaaa ggctcaagtg tccggccagg gcgacagcct ccacgagcac atcgccaacc 3661 tggccgggag ccccgctatt aagaagggaa tcctccagac cgtcaaggtg gttgacgagt 3721 tggtcaaggt gatgggcaga cataagccag agaacatcgt catcgagatg gccagggaaa 3781 accagaccac ccagaaggga cagaagaact cccgtgagcg catgaagagg attgaggagg 3841 gcatcaagga gcttggttcc cagatcctca aggagcaccc cgtggaaaac acccaactcc 3901 agaatgagaa gctgtatttg tactaccttc agaacgggcg cgatatgtac gttgaccaag 3961 agctggacat caaccgcctc agcgactacg acgtggatca cattgtccct cagtcctttc 4021 tcaaggacga ctctatcgac aacaaggtgt tgactagaag cgataagaac cgcggcaagt 4081 ccgacaacgt cccatccgag gaggtggtca agaagatgaa gaattactgg aggcagctgc 4141 ttaacgccaa gctcatcacc cagcgcaagt tcgacaactt gaccaaggcc gagcgtggag 4201 gcctcagcga actggacaag gctggattca tcaagaggca acttgttgaa acccgccaga 4261 ttaccaagca cgtggcccag atcctcgact cccgtatgaa cactaagtac gatgagaacg 4321 acaagctgat ccgcgaggtc aaagtgatta ccctcaagag caagctcgtg tctgacttca 4381 gaaaggactt ccaattctac aaggttaggg agatcaacaa ctaccaccac gcccacgatg 4441 cttaccttaa tgccgtggtc ggcaccgcct tgatcaagaa gtaccccaag ctggagtccg 4501 agttcgtgta tggtgactac aaggtctacg acgttcgcaa gatgatcgct aagagcgagc 4561 aggagattgg caaggccacc gccaagtact tcttctactc caacatcatg aacttcttta 4621 agaccgagat cactttggct aacggggaga tccgtaagcg ccctctcatt gaaaccaacg 4681 gagaaaccgg cgagatcgtg tgggacaagg gcagggactt cgccaccgtc agaaaagtgc 4741 tgagcatgcc ccaagtcaac atcgtgaaga aaaccgaggt tcagactgga ggcttctcca 4801 aggagtctat cctcccaaag cgcaattccg ataagttgat tgcccgtaag aaggactggg 4861 accccaagaa gtacggtgga ttcgacagcc caaccgtcgc ctactccgtg cttgtcgtgg 4921 ctaaagttga gaagggcaag agcaagaagc tcaagtccgt caaggagctg ctcgggatca 4981 ccatcatgga gcgcagctcc ttcgagaaga accctattga tttccttgag gccaagggct 5041 acaaggaagt gaagaaggac ttgatcatca agctccccaa gtactctctg ttcgagcttg 5101 agaacggaag gaagcgtatg ctcgcctccg ctggcgagct gcaaaaggga aacgagttgg 5161 ccctcccaag caagtacgtc aacttcctgt acctcgcctc ccactatgag aagctcaagg 5221 gcagccccga ggacaacgaa cagaagcagt tgtttgtgga gcagcataag cactaccttg 5281 acgagatcat tgagcagatc agcgagttct ccaagcgcgt tatcctggct gacgccaatc 5341 tcgataaagt cctttctgcc tacaacaagc acagggacaa gcctatcaga gagcaggctg 5401 aaaacattat ccacttgttc accctcacca acctgggtgc cccagccgcc ttcaagtact 5461 tcgacactac catcgaccgc aagcgttaca cctccaccaa ggaagtgctc gatgctaccc 5521 ttatccacca gagcattact gggttgtacg aaaccaggat cgacctgtcc caactcggcg 5581 gagactccgg cggatccggt ggctctggag ggtccaccaa cctctcagac atcattgaaa 5641 aggagactgg aaagcagctg gtcatccaag agagcatcct gatgcttccc gaggaggtgg 5701 aggaggttat cggaaacaag cccgagtcgg atattctcgt gcacaccgcc tacgacgagt 5761 ccacagacga gaatgtcatg ctcttgacca gcgatgctcc agaatacaag ccttgggcct 5821 tggtcatcca ggattccaat ggcgaaaaca agatcaaaat gctcagtggc ggttctggag 5881 gctccggtgg gagcacgaac ctttccgaca tcattgagaa ggagactggc aagcagttgg 5941 tgatccaaga gtcaatcctc atgctgccgg aggaagtcga ggaggtgatc ggcaacaagc 6001 ctgagagcga cattctcgtt cacaccgcat acgacgagtc gaccgacgag aacgtcatgc 6061 ttttgacctc cgacgctcca gagtacaagc cctgggccct ggtgatccag gactctaacg 6121 gcgagaacaa gatcaagatg ctcagcggag gctccaagcg ccccgccgcc accaagaagg 6181 ctggccaggc caagaagaag aagtaacccg ggccaggaac cgtaaaaaat gtgagcgagt 6241 aacaacctga tcttgagagc tattctagtc actcataaga tatctgcata ggaggtcagc 6301 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 6361 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 6421 ccgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 6481 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 6541 ccgtgttgac gccgggcaag agcaactcgg acaccgcata cactattctc agaatgactt 6601 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 6661 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 6721 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 6781 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 6841 gccagcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 6901 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 6961 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggctc 7021 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 7081 cacgacgggg agccaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 7141 ctcactgatt aagcattggt gatctcatgg ccaaaatccc ttaacgtgag ttttcgttcc 7201 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 7261 gcgtaatctg ctgcttgcaa acaaaaaaac catcgctacc agcggtggtt tgtttgccgg 7321 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 7381 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 7441 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 7501 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 7561 cggggggttc gtgctcacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 7621 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 7681 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 7741 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 7801 gctcgtcagg ggggcggagc ctatggaaa //