LOCUS       V043857                 7082 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043857
VERSION     V043857
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             962..1675
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1715..3412
                     /label="DONSON(NM_017613)"
                     /note="DONSON(NM_017613)"
                     /gene="DONSON"
     polyA_signal    3423..4045
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        4074..4403
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4442..4460)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4474..4490)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4498..4514
                     /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(4522..4552)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4567..4588
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4876..5464)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5635..6495)
                     /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(6496..6600)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6627..7055
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1141 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1381 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1441 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1561 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggcggc
     1681 cgcgggtgga ggcggatcag gaggcggtgg atctatggcc ctttcggtgc ccggctactc
     1741 accgggcttc cgaaagccgc ccgaggtagt gcggctccga cggaaaaggg cccggagccg
     1801 tggagctgcc gcctccccgc cccgtgagct gacggagccg gcggcccgcc gagccgccct
     1861 ggtggcgggg ctgcctcttc gccctttccc tgctgcgggg ggcagaggcg gtggcagcgg
     1921 cggcggcccg gccgctgctc ggaggaaccc cttcgcccgc ctggacaacc gaccgcgggt
     1981 cgccgcggag ccccccgacg ggccggcccg cgagcagccg gaggccccgg tcccgttttt
     2041 agattctaat caagaaaatg atttgctatg ggaagagaag tttcctgaaa gaacaactgt
     2101 tactgaatta cctcagactt cacatgtatc attctccgag cctgatattc cgtcctcaaa
     2161 aagtactgag ttacctgtgg actggagtat taaaacgcga ctccttttca cctcttctca
     2221 accctttacc tgggcagatc atttgaaagc acaggaagaa gctcaaggtc ttgtccagca
     2281 ttgtagggca acagaagtta ctttgcctaa aagtatacag gatcccaaac tctcctctga
     2341 gctccgttgt accttccagc agagccttat ctattggctc caccctgctt tgtcttggct
     2401 accactgttc cctcgtattg gagctgatag aaaaatggct ggaaagacaa gtccttggtc
     2461 aaatgatgca accctgcagc atgttttaat gagtgactgg tctgtgagct ttacttctct
     2521 atataatttg ctgaagacaa aactttgccc ctatttctac gtttgtacct atcagtttac
     2581 tgtcctgttc cgagcagcag gattagctgg aagtgactta atcacagctc tcatatctcc
     2641 aacaactcga ggtttaagag aagctatgag aaatgaaggt attgaatttt ctctgccttt
     2701 aataaaagaa agtggccata agaaggagac agcatctgga acaagcttgg gatatgggga
     2761 ggagcaagcc atcagtgatg aggatgaaga ggaaagtttt tcctggctgg aagagatggg
     2821 tgtgcaagat aaaattaaaa agccagacat actttctatc aagctgcgta aagagaaaca
     2881 tgaagtacaa atggatcaca gacctgaatc tgttgtgttg gtaaaaggaa tcaacacctt
     2941 tacattgctc aattttttga ttaactctaa gagtttagtt gctacctcag gtccacaggc
     3001 aggacttcct ccaaccctct tgtcccctgt tgctttccga ggtgccacaa tgcaaatgct
     3061 taaggcacgg agtgtgaatg tgaagacaca agctctttct ggatacagag accaatttag
     3121 tttggagatt acaggtccta tcatgcctca ttctctgcat tcactgacca tgctgctcaa
     3181 atcttcacag agtggatctt tctctgcagt actgtatcca cacgagccaa ctgctgtatt
     3241 taacatctgc ctgcaaatgg acaaagtact tgatatggag gttgttcata aggagcttac
     3301 taactgtggt ttgcacccta acactctgga gcaacttagt caaataccgt tacttgggaa
     3361 atcatcttta cggaatgtgg tgctgagaga ctacatttat aattggagat cctgaggatc
     3421 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     3481 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     3541 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     3601 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     3661 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     3721 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     3781 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     3841 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     3901 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     3961 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     4021 ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc
     4081 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4201 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4261 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4321 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4381 ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt
     4441 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     4501 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4561 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4621 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6601 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     6661 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6721 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6781 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6841 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6901 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6961 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     7021 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     7081 tt
//
