LOCUS       V036039                 8313 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036039
VERSION     V036039
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..3786
                     /label="DCBLD1(NM_001366458)"
                     /note="DCBLD1(NM_001366458)"
                     /gene="DCBLD1"
     CDS             3787..3804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3880..4104
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4150..4578
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4592..4921
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4988..5782
                     /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    5956..6077
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6126..6142)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6150..6166
                     /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(6174..6204)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6219..6240
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6528..7113)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7284..8144)
                     /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(8145..8249)
                     /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 catggtgccc ggcgcccgcg gcggcggcgc actggcgcgg
     1681 gctgccgggc ggggcctcct ggctttgctg ctcgcggtct ccgccccgct ccggctgcag
     1741 gcggaggagc tgggtgatgg ctgtggacac ctagtgactt atcaggatag tggcacaatg
     1801 acatctaaga attatcccgg gacctacccc aatcacactg tttgcgaaaa gacaattaca
     1861 gtaccaaagg ggaaaagact gattctgagg ttgggagatt tggatatcga atcccagacc
     1921 tgtgcttctg actatcttct cttcaccagc tcttcagatc aatatggtcc atactgtgga
     1981 agtatgactg ttcccaaaga actcttgttg aacacaagtg aagtaaccgt ccgctttgag
     2041 agtggatccc acatttctgg ccggggtttt ttgctgacct atgcgagcag cgaccatcca
     2101 gatttaataa catgtttgga acgagctagc cattatttga agacagaata cagcaaattc
     2161 tgcccagctg gttgtagaga cgtagcagga gacatttctg ggaatatggt agatggatat
     2221 agagatacct ctttattgtg caaagctgcc atccatgcag gaataattgc tgatgaacta
     2281 ggtggccaga tcagtgtgct tcagcgcaaa gggatcagtc gatatgaagg gattctggcc
     2341 aatggtgttc tttcgaggga tggttccctg tcagacaagc gatttctgtt tacctccaat
     2401 ggttgcagca gatccttgag ttttgaacct gacgggcaaa tcagagcttc ttcctcatgg
     2461 cagtcggtca atgagagtgg agaccaagtt cactggtctc ctggccaagc ccgacttcag
     2521 gaccaaggcc catcatgggc ttcgggcgac agtagcaaca accacaaacc acgagagtgg
     2581 ctggagatcg atttggggga gaaaaagaaa ataacaggaa ttaggaccac aggatctaca
     2641 cagtcgaact tcaactttta tgttaagagt tttgtgatga acttcaaaaa caataattct
     2701 aagtggaaga cctataaagg aattgtgaat aatgaagaaa aggtgtttca gggtaactct
     2761 aactttcggg acccagtgca aaacaatttc atccctccca tcgtggccag atatgtgcgg
     2821 gttgtccccc agacatggca ccagaggata gccttgaagg tggagctcat tggttgccag
     2881 attacacaag gtaatgattc attggtgtgg cgcaagacaa gtcaaagcac cagtgtttca
     2941 actaagaaag aagatgagac aatcacaagg cccatcccct cggaagaaac atccacagga
     3001 ataaacatta caacggtggc tattccattg gtgctccttg ttgtcctggt gtttgctgga
     3061 atggggatct ttgcagcctt tagaaagaag aagaagaaag gaagtccgta tggatcagca
     3121 gaggctcaga aaacagactg ttggaagcag attaaatatc cctttgccag acatcagtca
     3181 gctgagttta ccatcagcta tgataatgag aaggagatga cacaaaagtt agatctcatc
     3241 acaagtgata tggcagatta ccagcagccc ctcatgattg gcaccgggac agtcacgagg
     3301 aagggctcca ccttccggcc catggacacg gatgccgagg aggcaggggt gagcaccgat
     3361 gccggcggcc actatgactg cccgcagcgg gccggccgcc acgagtacgc gctgcccctg
     3421 gcgcccccgg agcccgagta cgccacgccc atcgtggagc ggcacgtgct gcgcgcccac
     3481 acgttctctg cgcagagcgg ctaccgcgtc ccagggcccc agcccggcca caaacactcc
     3541 ctctcctcgg gcggcttctc ccccgtagcg ggtgtgggcg cccaggacgg agactatcaa
     3601 aggccacaca gcgcacagcc tgcggacagg ggctacgacc ggcccaaagc tgtcagcgcc
     3661 ctcgccaccg aaagcgggca ccctgactct cagaagcccc caacgcatcc cgggacgagt
     3721 gacagctatt ctgcccccag agactgcctc acacccctca accagacggc catgactgcc
     3781 cttttgcacc accaccacca ccactaggat cctgatatcc agcacagtgg cggccgctcg
     3841 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3901 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3961 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4021 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4081 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     4141 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4201 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4261 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4321 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4381 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4441 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4501 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4561 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4621 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4741 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4861 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4921 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4981 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5041 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5101 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5161 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5221 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5281 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5341 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5401 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5461 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5521 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5581 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5641 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5701 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5761 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5821 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5881 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5941 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6061 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     7081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7561 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8281 gacggatcgg gagatctccc gatcccctat ggt
//
