LOCUS       V021498                 6710 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021498
VERSION     V021498
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             902..1498
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /note="also known as hA3A"
                     /product="human cytidine deaminase (C to U editing enzyme)
                     that efficiently edits methylated cytosines (Wang et al.,
                     2018)"
                     /protein_id=""
                     /transl_table=1
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
                     /label="APOBEC3A"
     CDS             1514..2233
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="cyan variant of GFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG
                     IKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="CFP"
     polyA_signal    2277..2501
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2547..2975
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2989..3318
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3385..4179
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4353..4474
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4523..4539)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4547..4563
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4571..4601)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4616..4637
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4925..5510)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5681..6541)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6542..6646)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 catggaagcc agcccagcat ccgggcccag acacttgatg gatccacaca tattcacttc
      961 caactttaac aatggcattg gaaggcataa gacctacctg tgctacgaag tggagcgcct
     1021 ggacaatggc acctcggtca agatggacca gcacaggggc tttctacaca accaggctaa
     1081 gaatcttctc tgtggctttt acggccgcca tgcggagctg cgcttcttgg acctggttcc
     1141 ttctttgcag ttggacccgg cccagatcta cagggtcact tggttcatct cctggagccc
     1201 ctgcttctcc tggggctgtg ccggggaagt gcgtgcgttc cttcaggaga acacacacgt
     1261 gagactgcgt atcttcgctg cccgcatcta tgattacgac cccctatata aggaggcact
     1321 gcaaatgctg cgggatgctg gggcccaagt ctccatcatg acctacgatg aatttaagca
     1381 ctgctgggac acctttgtgg accaccaggg atgtcccttc cagccctggg atggactaga
     1441 tgagcacagc caagccctga gtgggaggct gcgggccatt ctccagaatc agggaaacgg
     1501 aggaggggga agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1561 ggtcgagctg gacggcgacg taaacggcca caggttcagc gtgtccggcg agggcgaggg
     1621 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1681 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc
     1741 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1801 gcgtaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1861 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1921 catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga
     1981 caagcagaag aacggcatca aggcccactt caagatccgc cacaacatcg aggacggcag
     2041 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2101 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2161 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2221 gctgtacaag tagctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2281 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2341 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2401 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2461 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2521 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2581 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2641 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2701 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2761 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2821 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2881 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2941 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3001 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3061 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3121 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3241 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3301 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3361 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3421 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3481 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3541 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3601 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3661 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3721 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3781 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3841 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3901 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3961 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4021 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4081 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4141 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4201 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4261 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4321 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4381 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4441 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4501 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5341 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6661 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
