LOCUS       V032230                 7765 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032230
VERSION     V032230
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..2424
                     /label="HSPD1(NM_002156)"
                     /note="HSPD1(NM_002156)"
                     /gene="HSPD1"
     CDS             2431..2457
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2461..2487
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2491..2517
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2541..3129
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3195..3243
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3445..3873
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3887..4216
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4283..5077
                     /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    5253..5374
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5423..5439)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5447..5463
                     /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(5471..5501)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5516..5537
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5825..6413)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6584..7444)
                     /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(7445..7549)
                     /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 ccaccatgct tcggttaccc
      721 acagtctttc gccagatgag accggtgtcc agggtactgg ctcctcatct cactcgggct
      781 tatgccaaag atgtaaaatt tggtgcagat gcccgagcct taatgcttca aggtgtagac
      841 cttttagccg atgctgtggc cgttacaatg gggccaaagg gaagaacagt gattattgag
      901 cagagttggg gaagtcccaa agtaacaaaa gatggtgtga ctgttgcaaa gtcaattgac
      961 ttaaaagata aatacaaaaa cattggagct aaacttgttc aagatgttgc caataacaca
     1021 aatgaagaag ctggggatgg cactaccact gctactgtac tggcacgctc tatagccaag
     1081 gaaggcttcg agaagattag caaaggtgct aatccagtgg aaatcaggag aggtgtgatg
     1141 ttagctgttg atgctgtaat tgctgaactt aaaaagcagt ctaaacctgt gaccacccct
     1201 gaagaaattg cacaggttgc tacgatttct gcaaacggag acaaagaaat tggcaatatc
     1261 atctctgatg caatgaaaaa agttggaaga aagggtgtca tcacagtaaa ggatggaaaa
     1321 acactgaatg atgaattaga aattattgaa ggcatgaagt ttgatcgagg ctatatttct
     1381 ccatacttta ttaatacatc aaaaggtcag aaatgtgaat tccaggatgc ctatgttctg
     1441 ttgagtgaaa agaaaatttc tagtatccag tccattgtac ctgctcttga aattgccaat
     1501 gctcaccgta agcctttggt cataatcgct gaagatgttg atggagaagc tctaagtaca
     1561 ctcgtcttga ataggctaaa ggttggtctt caggttgtgg cagtcaaggc tccagggttt
     1621 ggtgacaata gaaagaacca gcttaaagat atggctattg ctactggtgg tgcagtgttt
     1681 ggagaagagg gattgaccct gaatcttgaa gacgttcagc ctcatgactt aggaaaagtt
     1741 ggagaggtca ttgtgaccaa agacgatgcc atgctcttaa aaggaaaagg tgacaaggct
     1801 caaattgaaa aacgtattca agaaatcatt gagcagttag atgtcacaac tagtgaatat
     1861 gaaaaggaaa aactgaatga acggcttgca aaactttcag atggagtggc tgtgctgaag
     1921 gttggtggga caagtgatgt tgaagtgaat gaaaagaaag acagagttac agatgccctt
     1981 aatgctacaa gagctgctgt tgaagaaggc attgttttgg gagggggttg tgccctcctt
     2041 cgatgcattc cagccttgga ctcattgact ccagctaatg aagatcaaaa aattggtata
     2101 gaaattatta aaagaacact caaaattcca gcaatgacca ttgctaagaa tgcaggtgtt
     2161 gaaggatctt tgatagttga gaaaattatg caaagttcct cagaagttgg ttatgatgct
     2221 atggctggag attttgtgaa tatggtggaa aaaggaatca ttgacccaac aaaggttgtg
     2281 agaactgctt tattggatgc tgctggtgtg gcctctctgt taactacagc agaagttgta
     2341 gtcacagaaa ttcctaaaga agagaaggac cctggaatgg gtgcaatggg tggaatggga
     2401 ggtggtatgg gaggtggcat gttcctcgag tacccatacg acgtacctga ttatgcaact
     2461 tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga ctacgcatag
     2521 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2581 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2641 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2701 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2761 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2821 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2881 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     2941 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3001 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3061 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3121 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3181 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3241 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3301 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3361 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3421 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3601 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3781 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3841 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3901 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3961 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4021 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4081 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4141 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4201 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4261 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4321 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4381 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4441 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4501 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4561 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4621 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4681 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4741 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4801 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4861 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4921 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4981 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5041 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5101 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5161 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5221 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5401 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6001 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6241 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7561 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     7621 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7681 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7741 caattgcatg aagaatctgc ttagg
//
