LOCUS       V032826                 8236 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032826
VERSION     V032826
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..3726
                     /label="ADGRF4(NM_153838)"
                     /note="ADGRF4(NM_153838)"
                     /gene="ADGRF4"
     polyA_signal    3803..4027
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4073..4501
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4515..4844
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4911..5705
                     /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    5879..6000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6049..6065)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6073..6089
                     /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(6097..6127)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6142..6163
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6451..7036)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7207..8067)
                     /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(8068..8172)
                     /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 catgaaaatg aagtcccagg caaccatgat ttgctgctta
     1681 gtgttctttc tgtccacaga atgttcccac tatagatcca agattcacct aaaagctgga
     1741 gataaacttc aaagccctga agggaaaccc aagactggaa ggatccaaga gaaatgcgaa
     1801 ggaccttgta tttcttcttc caactgcagc cagccctgtg ctaaggactt tcatggagaa
     1861 ataggattta catgtaatca aaaaaagtgg caaaaatcag ctgaaacatg tacaagcctt
     1921 tctgtggaaa aactctttaa ggactcaact ggtgcatctc gcctttctgt agcagcacca
     1981 tctatacctc tgcatattct agactttcga gctccagaga ccattgagag tgtagctcaa
     2041 ggaatccgta agaactgccc ctttgattat gcctgcatca ctgacatggt gaaatcatca
     2101 gaaacaacat ctggaaatat tgcatttata gtggagttat taaaaaatat ttctacagac
     2161 ttgtctgata atgttactcg agagaaaatg aagagctata gtgaagtggc caaccacatc
     2221 ctcgacacag cagccatttc aaactgggct ttcattccca acaaaaatgc cagctcggat
     2281 ttgttgcagt cagtgaattt gtttgccaga caactccaca tccacaataa ttctgagaac
     2341 attgtgaatg aactcttcat tcagacaaaa gggtttcaca tcaaccataa tacctcagag
     2401 aaaagcctca atttctccat gagcatgaac aataccacag aagatatctt aggaatggta
     2461 cagattccca ggcaagagct aaggaagctg tggccaaatg catcccaagc cattagcata
     2521 gctttcccaa ccttgggggc tatcctgaga gaagcccact tgcaaaatgt gagtcttccc
     2581 agacaggtaa atggtctggt gctatcagtg gttttaccag aaaggttgca agaaatcata
     2641 ctcaccttcg aaaagatcaa taaaacccgc aatgccagag cccagtgtgt tggctggcac
     2701 tccaagaaaa ggagatggga tgagaaagcg tgccaaatga tgttggatat caggaacgaa
     2761 gtgaaatgcc gctgtaacta caccagtgtg gtgatgtctt tttccattct catgtcctcc
     2821 aaatcgatga ccgacaaagt tctggactac atcacctgca ttgggctcag cgtctcaatc
     2881 ctaagcttgg ttctttgcct gatcattgaa gccacagtgt ggtcccgggt ggttgtgacg
     2941 gagatatcat acatgcgtca cgtgtgcatc gtgaatatag cagtgtccct tctgactgcc
     3001 aatgtgtggt ttatcatagg ctctcacttt aacattaagg cccaggacta caacatgtgt
     3061 gttgcagtga catttttcag ccactttttc tacctctctc tgtttttctg gatgctcttc
     3121 aaagcattgc tcatcattta tggaatattg gtcattttcc gtaggatgat gaagtcccga
     3181 atgatggtca ttggctttgc cattggctat gggtgcccat tgatcattgc tgtcactaca
     3241 gttgctatca cagagccaga gaaaggctac atgagacctg aggcctgttg gcttaactgg
     3301 gacaatacca aagccctttt agcatttgcc atcccggcgt tcgtcattgt ggctgtaaat
     3361 ctgattgtgg ttttggttgt tgctgtcaac actcagaggc cctctattgg cagttccaag
     3421 tctcaggatg tggtcataat tatgaggatc agcaaaaatg ttgccatcct cactccactg
     3481 ctgggactga cctggggttt tggaatagcc actctcatag aaggcacttc cttgacgttc
     3541 catataattt ttgccttgct caatgctttc cagggttttt tcatcctgct gtttggaacc
     3601 attatggatc acaagataag agatgctttg aggatgagga tgtcttcact gaaggggaaa
     3661 tcgagggcag ctgagaatgc atcactaggc ccaaccaatg gatctaaatt aatgaatcgt
     3721 caaggatgag gatcctgata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     3781 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3841 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3901 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3961 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4021 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4201 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4261 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4381 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4441 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4501 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4561 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4621 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4681 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4741 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4801 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4861 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4921 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4981 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5041 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5101 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5161 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5221 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5281 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5341 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5401 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5461 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5521 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5581 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5641 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5701 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5761 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5821 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6001 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6061 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6121 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6181 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6241 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6961 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     7021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8221 cccgatcccc tatggt
//
