LOCUS       V046496                 7785 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046496
VERSION     V046496
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2465
                     /label="TM7SF3(NM_016551)"
                     /note="TM7SF3(NM_016551)"
                     /gene="TM7SF3"
     CDS             2472..2537
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2561..3149
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3215..3263
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3465..3893
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3907..4236
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4303..5097
                     /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    5273..5394
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5443..5459)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5467..5483
                     /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(5491..5521)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5536..5557
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5845..6433)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6604..7464)
                     /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(7465..7569)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg gttcctgcag ctgctggtcg
      781 tagcggtgct ggcatccgaa caccgggtgg ctggtgcagc cgaggtcttc gggaattcca
      841 gcgagggtct tattgaattt tctgtgggga aatttagata cttcgagctc aataggccct
      901 ttccagagga agctattttg catgatattt caagcaatgt gacttttctt attttccaaa
      961 tacactcaca gtatcagaat acaactgttt ccttttctcc gactctcctt tccaattcct
     1021 cggaaacagg cactgccagt ggactggttt tcatccttag accagagcag agtacatgca
     1081 cttggtactt ggggacttca ggcatacagc ctgtccagaa tatggctatc ctactctcct
     1141 actcagaaag agatcctgtc cctggaggct gtaatttgga gttcgattta gatattgatc
     1201 ccaacattta cttggagtat aatttctttg aaacgactat caagtttgcc ccagcaaacc
     1261 taggctatgc gagaggcgta gatcccccac catgtgacgc tgggacagac caggactcca
     1321 ggtggaggtt gcagtatgat gtctatcagt attttctgcc tgagaatgac ctcactgagg
     1381 agatgttgct gaagcatctg cagaggatgg tcagtgtgcc ccaggtgaag gccagtgctc
     1441 tcaaggtggt taccctaaca gctaatgata agacaagtgt ttccttctcc tccctcccgg
     1501 gacaaggtgt catatacaat gtcattgttt gggacccgtt tctaaataca tctgctgcct
     1561 acattcctgc tcacacatac gcttgcagct ttgaggcagg agagggtagt tgtgcttccc
     1621 taggaagagt gtcttccaaa gtgttcttca ctctttttgc cctgcttggt ttcttcattt
     1681 gtttctttgg acacagattc tggaaaacag aattattctt cataggcttt atcatcatgg
     1741 gattcttctt ttatatactg attacaagac tgacacctat caagtatgat gtgaatctga
     1801 ttctgacagc tgtcactgga agcgtcggtg gaatgttctt ggtagctgtg tggtggcgat
     1861 ttggaatcct ctcgatctgc atgctctgtg ttggactagt gctggggttc ctcatctcgt
     1921 cagtgacttt ctttactcca ctgggaaacc taaagatttt tcatgatgat ggtgtattct
     1981 gggtcacttt ctcttgcata gctatcctca ttccagtagt tttcatgggc tgcctaagaa
     2041 tactgaacat actgacttgt ggagtcattg gctcctattc ggtggtttta gccattgaca
     2101 gttactggtc cacaagcctt tcctacatca ctttgaacgt actcaagaga gcgctcaaca
     2161 aggatttcca cagagctttc acaaatgtgc cttttcaaac taatgacttc attatcctgg
     2221 cagtatgggg catgctggct gtaagtggaa ttacgttaca gattcgaaga gagagaggac
     2281 gaccgttctt ccctccccac ccatacaagt tatggaagca agagagagag cgccgagtga
     2341 caaacattct ggaccctagc taccacattc ctccattgag agagaggctc tatggccgat
     2401 taacccagat taaagggctc ttccagaagg agcagccagc tggagagaga acgcctttgc
     2461 ttctgggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     2521 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     2581 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2641 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2701 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2761 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2821 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2881 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2941 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3001 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3061 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3121 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3181 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3241 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3301 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3361 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3421 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3481 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3541 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3601 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3661 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3721 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3781 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3841 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4261 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4801 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5101 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5161 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5221 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5281 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5341 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5401 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5581 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5641 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5761 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6361 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6421 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6481 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6541 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6661 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6721 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6781 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6841 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6901 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6961 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7021 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7081 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7141 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7201 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7261 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7321 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7381 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7441 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7501 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7561 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7621 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7681 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7741 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
