LOCUS       V038935                 8632 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038935
VERSION     V038935
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             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..4122
                     /label="CARNS1(NM_001394579)"
                     /note="CARNS1(NM_001394579)"
                     /gene="CARNS1"
     polyA_signal    4199..4423
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4469..4897
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4911..5240
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5307..6101
                     /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    6275..6396
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6445..6461)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6469..6485
                     /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(6493..6523)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6538..6559
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6847..7432)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7603..8463)
                     /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(8464..8568)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgctcctt tgcctgtccc ctgcttggct gatgaaggtg
     1681 ccagcacccg ggcagccggg tgaggcagcc ctgctagtct ccaaggctgt gagcttccac
     1741 cctgggggcc tgacattcct ggatgacttt gtccccccgc gccgtgccac ctactttttg
     1801 gcaggcctgg gcctggggcc tggccggggc cgagaggcag cagaactcgc ccgtgacctg
     1861 acctgcccca caggagcttc ggctgagctg gcccggctgc tggaggaccg gctgctgaca
     1921 aggcagttgc tggcccagca gggtggtgtg gctgtgccag caaccctggc tttcacctac
     1981 aagccgccgg ggctgctgcg gggaggggat gccagcctag ggctacggct ggtggagctg
     2041 agtggcaaag agggccagga gacgctggtg aaagaggaag tggaggcttt tctgcgctcc
     2101 gaggccctgg gtgatatcct gcaggtagct gtgaagctca gtggctggcg ctggcggggg
     2161 cggcaggcat ggcgtctgca cccgcgggca gagctgggtg cagtggtgga cacagtgctg
     2221 gcgctgctgg agaagctgga ggaggaggag agtgtcctgg tggaggctgt gtacccacct
     2281 gcccagctgc cctgctcaga tggtccttca cccggccccg gcctggccgt gcgaatctgt
     2341 gctgtggtgt gtcggacaca gggtgatagg ccactgctga gcaaggtggt gtgcggcgtg
     2401 ggccgcgggg accgccctct acggcaccac aactccctgc cgaggacgct ggaggtggcg
     2461 ctggcccagt gcggcctggg cgaggaggcg caggtggcgg ctgtgcggca gcgcgtcaag
     2521 gcggcggccg aggccgcgct ggccgccgtg ctggctctgg aggccggcct gagtgccgag
     2581 cagcgcggcg ggcgccgggc gcacacggac ttcctgggcg tggatttcgc gctgacagcg
     2641 gccggcggcg tgctgacccc agtggccctg gagctgaacg gcggcctgtg tctggaggcg
     2701 tgcggcgcgc tggaggggct gtgggccgcg ccgcggctgg ggccggcggc cgacgaggcg
     2761 gtggcggcgc cgctggtgga gaccatgctt cggcggtcgg cgcgctgcct catggaggga
     2821 aaacagctgc tggtggtcgg cgctggcggc gtcagcaaga agttcgtgtg ggaggcggcg
     2881 cgcgactacg ggctccagct gcacctcgtg gagtcagacc ccaaccactt tgcatcccag
     2941 ttggtacaga ccttcatcca ctttgacatg acagagcacc ggagggatga ggagaacgca
     3001 cggctgctgg cagagttggt gcgggctcgc ggcctcaagc tagatggctg cttctcctac
     3061 tgggatgact gcctggtgct cacagccctg ctctgccagg agctaggtct gccctgcagc
     3121 tccccagctg ccatgcgcct ggctaagcag aagagcctca cccagctgca cctgttgcac
     3181 caccatggcc caccctggcc tgcgccctcc ctccatgctg tgccctgctg cccactggag
     3241 agtgaggctg atgtggagag ggccgtgcac caggtacccc tgccaggtgt catgaagctg
     3301 gagttcgggg caggtgcagt gggtgtccgg ctggtagagg atgcgccaca gtgccatgag
     3361 cacttttccc ggattacccg agacttgcag ggcgaggccg accacccagg cattgggctg
     3421 ggctggggca atgccatgct gctgatggag tttgtggagg gcaccgagca cgacgtggac
     3481 ctggtgttgt ttggtgggcg gttgctggct gcctttgtct ccgacaatgg ccctacgagg
     3541 ctgcctggct tcactgagac ggcggcctgc atgcccaccg ggctggcacc agagcaggag
     3601 gcacagatgg ttcaggcagc cttccgctgt tgcctgggct gcgggttgct cgatggagtc
     3661 ttcaacgtgg agctcaagct gaccggggct gggcctcggc ttatcgagat caacccccgc
     3721 atgggtggct tctacctgcg tgattggatc ctggagctct atggtgttga cctgctgctg
     3781 gctgctgtta tggtggcctg tggcttgcgt cctgccctgc ccacccgccc acgtgctcgt
     3841 ggccatctgg tgggcgtcat gtgccttgtg tcccagcacc tgcaggccct gagttccacc
     3901 gccagccgtg agaccctgca ggccctgcac gaccgtggac tgctacgcct caatctgctg
     3961 gaggaggccc tggtgcctgg cgagtatgag gagccctact gcagtgtggc ctgtgccgga
     4021 cccagcccca ccgaggcccg tctccgcctg ctgggcctct gccagggcct gggcatcgat
     4081 gggcccagct accctgttgc ccacttcctg tctcacttca aatagggatc ctgatatcca
     4141 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     4201 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4261 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4321 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4381 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4441 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4921 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     5281 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5341 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5401 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5461 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5521 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5581 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5641 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5701 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5761 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5821 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5881 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5941 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6001 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6061 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6121 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6181 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6241 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6421 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7381 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8581 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
