LOCUS       V018997                 7301 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018997
VERSION     V018997
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             671..1981
                     /label="TMPRSS4(NM_019894)"
                     /note="TMPRSS4(NM_019894)"
                     /gene="TMPRSS4"
     CDS             1988..2053
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2077..2665
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2731..2779
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2981..3409
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3423..3752
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3819..4613
                     /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    4789..4910
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4959..4975)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4983..4999
                     /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(5007..5037)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5052..5073
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5361..5949)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6120..6980)
                     /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(6981..7085)
                     /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 taccgccacc atgttacagg atcctgacag tgatcaacct ctgaacagcc tcgatgtcaa
      721 acccctgcgc aaaccccgta tccccatgga gaccttcaga aaggtgggga tccccatcat
      781 catagcacta ctgagcctgg cgagtatcat cattgtggtt gtcctcatca aggtgattct
      841 ggataaatac tacttcctct gcgggcagcc tctccacttc atcccgagga agcagctgtg
      901 tgacggagag ctggactgtc ccttggggga ggacgaggag cactgtgtca agagcttccc
      961 cgaagggcct gcagtggcag tccgcctctc caaggaccga tccacactgc aggtgctgga
     1021 ctcggccaca gggaactggt tctctgcctg tttcgacaac ttcacagaag ctctcgctga
     1081 gacagcctgt aggcagatgg gctacagcag caaacccact ttcagagctg tggagattgg
     1141 cccagaccag gatctggatg ttgttgaaat cacagaaaac agccaggagc ttcgcatgcg
     1201 gaactcaagt gggccctgtc tctcaggctc cctggtctcc ctgcactgtc ttgcctgtgg
     1261 gaagagcctg aagacccccc gtgtggtggg tgtggaggag gcctctgtgg attcttggcc
     1321 ttggcaggtc agcatccagt acgacaaaca gcacgtctgt ggagggagca tcctggaccc
     1381 ccactgggtc ctcacggcag cccactgctt caggaaacat accgatgtgt tcaactggaa
     1441 ggtgcgggca ggctcagaca aactgggcag cttcccatcc ctggctgtgg ccaagatcat
     1501 catcattgaa ttcaacccca tgtaccccaa agacaatgac atcgccctca tgaagctgca
     1561 gttcccactc actttctcag gcacagtcag gcccatctgt ctgcccttct ttgatgagga
     1621 gctcactcca gccaccccac tctggatcat tggatggggc tttacgaagc agaatggagg
     1681 gaagatgtct gacatactgc tgcaggcgtc agtccaggtc attgacagca cacggtgcaa
     1741 tgcagacgat gcgtaccagg gggaagtcac cgagaagatg atgtgtgcag gcatcccgga
     1801 agggggtgtg gacacctgcc agggtgacag tggtgggccc ctgatgtacc aatctgacca
     1861 gtggcatgtg gtgggcatcg ttagttgggg ctatggctgc gggggcccga gcaccccagg
     1921 agtatacacc aaggtctcag cctatctcaa ctggatctac aatgtctgga aggctgagct
     1981 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     2041 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     2101 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2161 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2221 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2281 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2341 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2401 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2461 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     2521 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2581 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2641 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     2701 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     2761 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     2821 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     2881 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     2941 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3001 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3061 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3121 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3181 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3241 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3301 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     3361 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3421 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3481 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3661 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3721 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3781 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4681 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4741 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4801 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4861 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4921 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4981 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5041 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5101 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5161 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5221 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5281 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5521 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5761 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6421 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6481 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6541 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6601 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6661 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6721 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6781 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6841 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6901 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6961 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7021 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7081 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7141 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     7201 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7261 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
