LOCUS       V045140                 7731 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045140
VERSION     V045140
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..2411
                     /label="Has2(NM_008216)"
                     /note="Has2(NM_008216)"
                     /gene="Has2"
     CDS             2418..2483
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2507..3095
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3161..3209
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3411..3839
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3853..4182
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4249..5043
                     /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    5219..5340
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5389..5405)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5413..5429
                     /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(5437..5467)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5482..5503
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5791..6379)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6550..7410)
                     /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(7411..7515)
                     /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 ccaccatgca ttgtgagagg tttctatgtg
      781 tcctgagaat aattggaact acactttttg gagtgtctct cctcctcgga atcacagctg
      841 cttatattgt tggctaccag tttatccaaa cagataatta ctacttctca tttggactgt
      901 acggtgcctt tttagcctcg catctcatca tccaaagcct ctttgccttt ttggaacacc
      961 ggaaaatgaa gaagtccctt gaaaccccga ttaaattgaa caaaacggta gcactctgca
     1021 tcgctgcgta ccaagaggac cctgactact tacggaaatg tttgcaatct gtgaaaaggc
     1081 tgacctaccc tgggattaaa gtcgtgatgg tcatcgatgg gaactcagac gacgaccttt
     1141 acatgatgga catattcagc gaagttatgg gcagggacaa atcggccacg tacatctgga
     1201 agaacaactt tcatgaaaag ggacctggtg agacagaaga gtcccataaa gaaagttcac
     1261 aacatgtcac ccaattggtc ttgtctaaca aaagtatttg catcatgcaa aaatggggtg
     1321 gaaagagaga agtcatgtac acagccttca gagcactggg gcgaagcgtg gattatgtac
     1381 aggtgtgtga ctcagatact atgcttgacc ctgcctcatc tgtggagatg gtgaaggtct
     1441 tagaggaaga ccctatggtt ggaggtgttg gaggagatgt ccagatttta aacaagtatg
     1501 attcctggat ctccttcctc agcagcgtga gatactggat ggcttttaat atagaaaggg
     1561 cctgccagtc ttattttggc tgtgtccagt gcataagcgg tcctctggga atgtacagaa
     1621 actccttgct gcatgaattt gtggaagact ggtacaatca ggaattcatg ggtaaccaat
     1681 gcagttttgg tgacgacagg caccttacca acagggtgtt gagtctgggc tatgcaacta
     1741 aatacacggc tcggtccaag tgccttactg aaactcccat agaatatctg agatggctga
     1801 accagcagac ccgttggagc aagtcctact tccgagagtg gctgtacaat gccatgtggt
     1861 ttcacaagca tcacttgtgg atgacctatg aagctgttat cactggattc tttcctttct
     1921 ttctcattgc cacagtcatc cagctcttct acaggggtaa aatctggaac atcctcctct
     1981 tcctgttaac tgtccagcta gtgggtctca tcaagtcatc ttttgccagc tgccttagag
     2041 gaaatatcgt catggtattc atgtctctgt attcagtgtt atacatgtca agtctacttc
     2101 ctgccaagat gtttgcaatt gcaaccataa acaaagctgg gtggggcaca tctggaagga
     2161 agaccattgt tgttaatttc ataggactta ttccagtgtc cgtgtggttt acaatccttc
     2221 taggtggtgt aattttcacc atttataagg aatctaaaaa gccattttcc gaatccaaac
     2281 agactgttct catcgtggga actttgatct atgcatgcta ctgggtcatg cttttgactc
     2341 tctatgtggt tctcatcaat aagtgtggca ggcggaagaa gggacaacag tatgacatgg
     2401 tgcttgatgt aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2461 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2521 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2581 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2641 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2701 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2761 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2821 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2881 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2941 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3001 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3061 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3121 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3181 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3241 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3301 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3361 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3421 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3481 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3541 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3601 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3661 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3721 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3781 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3841 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4021 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4081 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4141 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4201 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4261 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4321 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4381 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4441 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4501 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4561 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4621 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4681 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4741 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4801 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4861 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4921 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4981 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5041 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5101 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5161 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5221 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5281 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5341 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5401 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5461 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7561 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7621 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7681 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
