LOCUS       V018786                 7445 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018786
VERSION     V018786
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2125
                     /label="HTR2B(NM_000867)"
                     /note="HTR2B(NM_000867)"
                     /gene="HTR2B"
     CDS             2132..2197
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2221..2809
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2875..2923
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3125..3553
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3567..3896
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3963..4757
                     /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    4933..5054
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5103..5119)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5127..5143
                     /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(5151..5181)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5196..5217
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5505..6093)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6264..7124)
                     /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(7125..7229)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggctct ctcttacaga gtgtctgaac ttcaaagcac
      721 aattcctgag cacattttgc agagcacctt tgttcacgtt atctcttcta actggtctgg
      781 attacagaca gaatcaatac cagaggaaat gaaacagatt gttgaggaac agggaaataa
      841 actgcactgg gcagctcttc tgatactcat ggtgataata cccacaattg gtggaaatac
      901 ccttgttatt ctggctgttt cactggagaa gaagctgcag tatgctacta attactttct
      961 aatgtccttg gcggtggctg atttgctggt tggattgttt gtgatgccaa ttgccctctt
     1021 gacaataatg tttgaggcta tgtggcccct cccacttgtt ctatgtcctg cctggttatt
     1081 tcttgacgtt ctcttttcaa ccgcatccat catgcatctc tgtgccattt cagtggatcg
     1141 ttacatagcc atcaaaaagc caatccaggc caatcaatat aactcacggg ctacagcatt
     1201 catcaagatt acagtggtgt ggttaatttc aataggcatt gccattccag tccctattaa
     1261 agggatagag actgatgtgg acaacccaaa caatatcact tgtgtgctga caaaggaacg
     1321 ttttggcgat ttcatgctct ttggctcact ggctgccttc ttcacacctc ttgcaattat
     1381 gattgtcacc tactttctca ctatccatgc tttacagaag aaggcttact tagtcaaaaa
     1441 caagccacct caacgcctaa catggttgac tgtgtctaca gttttccaaa gggatgaaac
     1501 accttgctcg tcaccggaaa aggtggcaat gctggatggt tctcgaaagg acaaggctct
     1561 gcccaactca ggtgatgaaa cacttatgcg aagaacatcc acaattggga aaaagtcagt
     1621 gcagaccatt tccaacgaac agagagcctc aaaggtccta gggattgtgt ttttcctctt
     1681 tttgcttatg tggtgtccct tctttattac aaatataact ttagttttat gtgattcctg
     1741 taaccaaact actctccaaa tgctcctgga gatatttgtg tggataggct atgtttcctc
     1801 aggagtgaat cctttggtct acaccctctt caataagaca tttcgggatg catttggccg
     1861 atatatcacc tgcaattacc gggccacaaa gtcagtaaaa actctcagaa aacgctccag
     1921 taagatctac ttccggaatc caatggcaga gaactctaag tttttcaaga aacatggaat
     1981 tcgaaatggg attaaccctg ccatgtacca gagtccaatg aggctccgaa gttcaaccat
     2041 tcagtcttca tcaatcattc tactagatac gcttctcctc actgaaaatg aaggtgacaa
     2101 aactgaagag caagttagtt atgtactcga ggactacaaa gaccatgacg gtgattataa
     2161 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     2221 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2281 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2341 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2401 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2461 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2521 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2581 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     2641 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2701 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2761 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     2821 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     2881 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     2941 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3001 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3061 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     3121 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3481 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3541 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     4801 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4861 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4921 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5041 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     5701 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5941 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7261 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7321 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7381 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7441 ttagg
//
