LOCUS       V038658                 7924 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038658
VERSION     V038658
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..3414
                     /label="GBP2(NM_004120)"
                     /note="GBP2(NM_004120)"
                     /gene="GBP2"
     polyA_signal    3491..3715
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3761..4189
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4203..4532
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4599..5393
                     /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    5567..5688
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5737..5753)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5761..5777
                     /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(5785..5815)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5830..5851
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6139..6724)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6895..7755)
                     /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(7756..7860)
                     /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 catggctcca gagatcaact tgccgggccc aatgagcctc
     1681 attgataaca ctaaagggca gctggtggtg aatccagaag ctctgaagat cctatctgca
     1741 attacgcagc ctgtggtggt ggtggcgatt gtgggcctct atcgcacagg caaatcctac
     1801 ctgatgaaca agctggctgg gaagaaaaac ggcttctctc taggctccac agtgaagtct
     1861 cacaccaagg gaatctggat gtggtgtgtg cctcatccca agaagccaga acacacccta
     1921 gttctgctcg acactgaggg cctgggagat atagagaagg gtgacaatga gaatgactcc
     1981 tggatctttg ccttggccat cctcctgagc agcaccttcg tgtacaatag catgggaacc
     2041 atcaaccagc aggccatgga ccaacttcac tatgtgacag agctgacaga tcgaatcaag
     2101 gcaaactcct cacctggtaa caattctgta gacgactcag ctgactttgt gagctttttt
     2161 ccagcatttg tgtggactct cagagatttc accctggaac tggaagtaga tggagaaccc
     2221 atcactgctg atgactactt ggagctttcg ctaaagctaa gaaaaggtac tgataagaaa
     2281 agtaaaagct ttaatgatcc tcggttgtgc atccgaaagt tcttccccaa gaggaagtgc
     2341 ttcgtcttcg attggcccgc tcctaagaag taccttgctc acctagagca gctaaaggag
     2401 gaagagctga accctgattt catagaacaa gttgcagaat tttgttccta catcctcagc
     2461 cattccaatg tcaagactct ttcaggtggc attccagtca atgggcctcg tctagagagc
     2521 ctggtgctga cctacgtcaa tgccatcagc agtggggatc taccctgcat ggagaacgca
     2581 gtcctggcct tggcccagat agagaactca gccgcagtgg aaaaggctat tgcccactat
     2641 gaacagcaga tgggccagaa ggtgcagctg cccacggaaa ccctccagga gctgctggac
     2701 ctgcacaggg acagtgagag agaggccatt gaagtcttca tgaagaactc tttcaaggat
     2761 gtggaccaaa tgttccagag gaaattaggg gcccagttgg aagcaaggcg agatgacttt
     2821 tgtaagcaga attccaaagc atcatcagat tgttgcatgg ctttacttca ggatatattt
     2881 ggccctttag aagaagatgt caagcaggga acattttcta aaccaggagg ttaccgtctc
     2941 tttactcaga agctgcagga gctgaagaat aagtactacc aggtgccaag gaaggggata
     3001 caggccaaag aggtgctgaa aaaatatttg gagtccaagg aggatgtggc tgatgcactt
     3061 ctacagactg atcagtcact ctcagaaaag gaaaaagcga ttgaagtgga acgtataaag
     3121 gctgaatctg cagaagctgc aaagaaaatg ttggaggaaa tacaaaagaa gaatgaggag
     3181 atgatggaac agaaagagaa gagttatcag gaacatgtga aacaattgac tgagaagatg
     3241 gagagggaca gggcccagtt aatggcagag caagagaaga ccctcgctct taaacttcag
     3301 gaacaggaac gccttctcaa ggagggattc gagaatgaga gcaagagact tcaaaaagac
     3361 atatgggata tccagatgag aagcaaatca ttggagccaa tatgtaacat actctaagga
     3421 tcctgatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     3481 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3541 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3601 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3661 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3721 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3781 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3841 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3901 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3961 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4021 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4081 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4141 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4201 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4381 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4441 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4501 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4561 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5101 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5401 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5581 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5701 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5761 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5821 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5881 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6661 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7861 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7921 tggt
//
