LOCUS       V031567                 8397 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031567
VERSION     V031567
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..3203
                     /label="Git2(NM_001347400)"
                     /note="Git2(NM_001347400)"
                     /gene="Git2"
     CDS             3240..3959
                     /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"
     polyA_signal    3997..4221
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4267..4695
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4709..5038
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5105..5899
                     /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    6073..6194
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6243..6259)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6267..6283
                     /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(6291..6321)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6336..6357
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6645..7230)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7401..8261)
                     /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(8262..8366)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgt cgaagcggct ccggagcagc gacgtgtgcg
      961 ccgactgcaa cgggcctgat ccttcctggg cgtcagtaaa taggggaact ttcatatgtg
     1021 acgaatgttg cagtgtccat cggagcctgg ggcgccatat ctcccaagtg aggcatctta
     1081 aacacacagc atggcctcca accctgctcc agatggttga gaccttgtac aacaatggtg
     1141 ctaactctat atgggaacac tctctgctgg accctgcctc tatcatgagt gggaggcgga
     1201 aagctaaccc acaggacaaa gttcacccca ataaagcaga attcatcaga gccaagtatc
     1261 agatgttagc gttcgtccac cgattgccct gccgcgagga cgacagcgtg acggccaaag
     1321 atctcagtaa gcaactccat tcaagtgtga gaacagggaa tcttgagacc tgtctgagac
     1381 tgttgtcttt aggagcccaa gccaacttct ttcatcctga aaaaggaagc actccactcc
     1441 acgtcgcctc caaagcagga cagatcttac aggctgaatt gctggccgtg tatggagctg
     1501 acccaggcac acaagattct agtgggaaga ctcctgttga ctatgcaagg caaggagggc
     1561 accatgagtt ggcagaacgc ctcatagaaa tacagtacga gctgacggat agactggcct
     1621 tctacctctg cggccggaaa ccagatcata aaagcggaca gcactttcta attcctcaga
     1681 gggcagacag cagcctggat ttgtctgagt tggcaaaagc tgcgaagaag aaacttcaat
     1741 ctctgagtaa tcatttgttt gaagaacttg ccatggatgt gtacgacgaa gttgacaggc
     1801 gagagactga cgcagtctgg cttgccacgc agaaccacag taccttggta accgagacga
     1861 ccgtcgtccc tttccttccg gtcaaccctg agtactcctc cacacgaaac cagggcagac
     1921 agaaattagc tcggtttaac gcccacgagt ttgctacgct ggtcatcgac atcctcagtg
     1981 atgccaagag gagacaacag ggcagtcctc tgtccaggtc aaaagacaat gtggagctca
     2041 tattgagaac agtcagtacc cagcacagca ctgagagcca agacaacgac cagccagact
     2101 atgacagtgt ggcctccgat gaagacacag acgtggagac gagggcaagc aaggcaaacc
     2161 ggcagaagag cctagattca gatttgtcag atggaccagt cactgtgcag gaatttatgg
     2221 aggtcaaaag cgccctagtg gcttctgagg ccaagagaca gcagctaatg aaggtgaata
     2281 acaacttgag tgacgagctg agaattatgc agaaaaagct tcaaactctc cagagtgaga
     2341 actcaagcct caggagacag gccacagcca gtgcctgtca ggtgcagact ggctctgacc
     2401 acaaggacac tgcaagccac tcgtccctaa agcgacggcc atctgcccgg ggcagtagac
     2461 ccatgtccat gtacgagact ggatcaggcc agaagccata cctcccaatg ggagaagcca
     2521 gccaccctga ggagagccgg acgagacttc aacccttccc cacgcatatt gggaggagtg
     2581 cacttgtgac ctcctcttcg tctctgccct ccttcccctc cacactttcc tggtccaggg
     2641 acgaaagcgc ccgaagggcc tccagactgg agaagcagaa cagcaccccg gagagtgatt
     2701 atgacaacac tgcctgtgac cctgagccag acgacacggg gtcaacccgg aagggacggc
     2761 agcggagcat gctctggcaa ggtgatggct tattgccaga tacagcagaa ccccactcgg
     2821 tccccagccc caccctcccc agcacagaag acgtcatccg gaagaccgag cagatcacca
     2881 aaaacatcca ggagctcctg cgagcagccc aggagaacaa gcatgacagc tatattccct
     2941 gctcagagag gatacacgtt gctgtaaccg agatggcagc cctgttcccc aaaaaaccca
     3001 agtctgacac ggtgaggact tccctccgcc tactcacgtc cagtgcctac cggctccagt
     3061 cggagtgcag aaaggctctc ccaggagact ccagcttacc cacagatgtg cagctggtca
     3121 cacagcaggt catccagtgt gcctatgaca tcgccaaggc cgccaaacag ctggtcacaa
     3181 tcaccaccaa agagaacagc agctcgagtg gcggtggagg aagcggcgga ggtggatcta
     3241 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3301 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3361 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3421 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3481 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3541 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3601 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3661 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3721 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3781 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3841 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3901 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtagt
     3961 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     4021 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4081 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4141 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4201 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4261 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4321 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4381 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4441 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4501 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4561 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4621 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4681 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4921 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4981 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5041 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     5101 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5161 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5221 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5281 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5341 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5401 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5461 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5521 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5581 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5641 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5701 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5761 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5821 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5881 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5941 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6001 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6061 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6181 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6241 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6301 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6361 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
