LOCUS       V045353                 8685 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045353
VERSION     V045353
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..3365
                     /label="Hps3(NM_001146323)"
                     /note="Hps3(NM_001146323)"
                     /gene="Hps3"
     CDS             3372..3437
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3461..4049
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4115..4163
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4365..4793
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4807..5136
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5203..5997
                     /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    6173..6294
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6343..6359)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6367..6383
                     /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(6391..6421)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6436..6457
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6745..7333)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7504..8364)
                     /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(8365..8469)
                     /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 ccaccatggt gcgtctgtac aacctgcacc
      781 cgtttgggtc gcagcaggta gtcccctgcc agtgggagcc ggagcaggtc tgctgcggcg
      841 ggagcgacgc gctgttcgtg gctgccggct gcaaggtgga ggcgttcgct gttcagggcg
      901 aagagctgtg ccggcagcgc tgcgccttct ccacgctagg cagggtgctg cgcatggcct
      961 acagcgaagc cgaagtctta ctcctaaaac tggagtcaga ccctatacat ggagagagtg
     1021 ttgaccacca cccacaagaa accagcaatc cattgaaaga ggcggaaggt gtcagtaatg
     1081 aaacttcaca acttgagtca gaagattttg ttatctgtct aaaacccatg gaactcattg
     1141 gtgaaaaatg tgaacagtct gggatatctg ttaaactgga gtccacagga ttagaagatg
     1201 aaaaagtaaa atatttacgt gttcggcatc tattgtatag acgttttgct ccagatattt
     1261 cgtcctatgt cttgtctgat aacatcaagt tacattccct tcagctgcta cccattcacc
     1321 aaagtggttt tcatcctgat gaaaatgatt tatcgccgaa aaaagagatg ccaaatctct
     1381 tctgtttctt ctcgttacct catgtgggct atctgtacat ggttgtgaag tctgttgaat
     1441 taatgtcagt ctattggtat cctgagaagt ctcaacaagc cgtgctcaca ccacagttct
     1501 tgcatgtcat cacaagccaa agcctgcagt gtttcacggt gcgatgcagt gcagcggtgg
     1561 cacatgagga agacctatac atggatacta cgctaaaggc ttgcccacct gtcagtatgg
     1621 acgtttgtgc tctaaggata cagcttttca taggcttgaa agccatctgt cactttaaaa
     1681 accacattat acttctgacg aaggcagagc ctgaggccat tccagagaga agagagtcac
     1741 ccaagaaact tatttctaga aaagatgcca gtgttaggag cggaacccct cacgtggctg
     1801 aagcggcatg gaatctgtat ctcgtgaaca ccacggcacc agtgcagctg tacaaagaaa
     1861 tggtagacta tagtaacagc tacaagaccg tgaaaaccga gagctgtctc cacctgctca
     1921 gtgaggctca tctgctagtg agagctgccc tgatggatgg cagtcagctg gagcctgcag
     1981 agaaagcaga actcctggaa gcttttaaag aaagctgtgg acaccttgga gactgttaca
     2041 gcaggcttac cactgagcaa tcccatctcg ccttgccgta ctataagatg tctggtttgt
     2101 ctctggctga agtcttggcc cgtgtggact ggacagaaga gagtgaatca cagaaatatg
     2161 agagaggact agtattttat attaatcatt ctctttatga aaacctggat gaagaattaa
     2221 gtaaagaatt agcagcaaaa gtggctcaga tattccacat ggctgagccg aagcagctgc
     2281 ctcatgttct ctctagccct tctatgaaga atattgatcc cttaactgcc ttgcattatt
     2341 taaggaagct ggatagttgt ggagtttcac ccgtcttagt gaccttgacc aaggctgcag
     2401 tggccttgaa aatgggagat cttgacatgt acagaaatga gatgaaaagc cattcggaga
     2461 tgaagctggt gtatggcttc attctggaac cccggctgtt aattcaacag tggaaaggac
     2521 agattgttcc tacagaactt gcgattgact tgaaggaaac tcagcctgga ttgcttgtgg
     2581 cttcagtcct gggcttacag aagaatgata aaattggaat tgtagaaaca gattctttct
     2641 ttaaggtgct gtgtggaaag gatgaagacg cagttccgca gcttttgata gacttctggg
     2701 aagctcagct cgtagcttgt ctcccaaatg tggtacttga ggaactcttt ttcaagctca
     2761 tctctcagta cgtttggagg ttatctgaga ggcggtgtcc tgacaccgta ccactgagaa
     2821 cagcggagga tttgataaat gcctgcagtc attatggcct agtaaatcca tgggttcatg
     2881 tgttaacgac atctgattcc ttagctgata aaaattacac agacgacctt ctgaaattac
     2941 agtctctcat ctgtagccct tcattggacg tagcctccat tattccgttt ttggagccac
     3001 tctcagaaga cactgttgcc ggcctcagta ctcatgctct ctgccataca cgcttacaag
     3061 agtatgaaca gtgcatagac acgttgctag agagatgccc agaggccgtc attgcgtatg
     3121 ctaatcagga actgaaggaa gatcactgga ttctgtggtg gaaaaagctg ctgcctgagc
     3181 tctgccagag agtcaagagt ggtggcgaga ggtcccacct tcacctgtca ttgttaaaag
     3241 aaaccttgtc ggttattgct gtgggactgg acctgaggga tttcttgaat gttctcccag
     3301 aagatggggc agcagcattt ttcctgccct acctcctttt ctgcagccgg aagaaatcac
     3361 taactggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3421 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3481 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3541 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3601 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3661 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3721 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3781 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3841 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3901 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3961 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4021 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4081 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4141 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4201 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4261 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4321 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4501 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4621 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4681 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4741 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4801 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4861 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4921 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4981 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5041 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5101 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5161 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5221 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5281 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5341 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5401 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5461 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5521 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5581 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5641 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5701 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5761 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5821 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5881 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5941 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6001 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6061 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6121 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6181 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6241 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6301 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6361 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6421 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6481 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6541 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6721 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7381 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7441 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7501 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7621 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7801 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8341 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8461 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8521 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8581 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8641 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
