LOCUS       V045722                 6931 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045722
VERSION     V045722
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             706..1590
                     /label="Vdac2(NM_011695)"
                     /note="Vdac2(NM_011695)"
                     /gene="Vdac2"
     CDS             1597..1623
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1627..1653
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1657..1683
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    1707..2295
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2361..2409
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2611..3039
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3053..3382
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3449..4243
                     /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    4419..4540
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4589..4605)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4613..4629
                     /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(4637..4667)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4682..4703
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4991..5579)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5750..6610)
                     /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(6611..6715)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc tgagtgctgt
      721 gtaccggtat gcccacggcc gatgtgtatc cctccaccct atgctgacct cggcaaagct
      781 gccagagaca ttttcaacaa aggatttggc tttgggctgg tgaagctgga tgtgaagacg
      841 aagtcatgca gcggtgtgga attttcaaca tctggctcat ctaatacaga cactggtaaa
      901 gttagcggga ccttggagac caagtacaaa tggtgtgagt atggtctgac tttcacagag
      961 aagtggaaca ccgataacac tctggggaca gagattgcaa ttgaagacca gatttgtcaa
     1021 ggtttgaaac tgacttttga caccaccttt tcaccgaaca caggaaagaa aagtggtaaa
     1081 atcaagtctg cttacaagag ggagtgtata aacctcggct gtgatgttga ctttgatttt
     1141 gctggacctg ccatccatgg gtcagctgtc tttggttacg agggctggct tgctgggtac
     1201 caaatgacct ttgacagtgc caagtcaaag ctgacaagga gtaactttgc agtcggctac
     1261 aggactgggg acttccagct acacacaaat gtaaataatg ggacagaatt tggaggatca
     1321 atttatcaga aagtatgtga agattttgac acttcagtaa acctcgcttg gacatcaggt
     1381 accaactgca ctcgttttgg cattgcagct aaataccagt tggatcctac tgcttctatc
     1441 tctgcaaagg tcaacaactc tagtttaatt ggagtgggct atactcagac tctgaggcct
     1501 ggtgtgaagc ttacactgtc tgctctggta gacgggaaga gctttaatgc tggaggccac
     1561 aaacttgggc ttgccttgga attggaggct ctcgagtacc catacgacgt acctgattat
     1621 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     1681 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     1741 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1801 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1861 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     1921 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     1981 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2101 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2161 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2221 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2281 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     2341 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     2401 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     2461 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     2521 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     2581 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2641 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2701 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2761 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2821 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2881 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2941 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3001 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3061 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3121 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3181 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3241 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3301 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3361 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3421 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3481 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3541 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3601 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3661 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3721 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3781 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3841 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3901 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3961 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4021 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4081 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4141 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4201 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4261 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4321 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4381 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4441 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4501 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4561 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4621 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4681 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4741 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4801 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4861 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4921 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4981 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5041 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5101 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5161 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5221 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5281 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5341 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5401 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5461 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5521 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5821 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5881 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5941 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6001 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6061 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6121 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6181 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6241 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6301 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6361 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6421 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6481 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6541 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6601 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6661 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6721 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     6781 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6841 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     6901 gaccgacaat tgcatgaaga atctgcttag g
//
