LOCUS       V025743                 8157 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025743
VERSION     V025743
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..2837
                     /label="Polh(NM_030715)"
                     /note="Polh(NM_030715)"
                     /gene="Polh"
     CDS             2844..2909
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2933..3521
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3587..3635
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3837..4265
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4279..4608
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4675..5469
                     /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    5645..5766
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5815..5831)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5839..5855
                     /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(5863..5893)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5908..5929
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6217..6805)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6976..7836)
                     /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(7837..7941)
                     /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 ccaccatggc tcctgggcag aatcgagtgg
      781 ttgctcttgt agacatggac tgcttttttg tacaagtgga acagcggcaa aatcctcatt
      841 tgaggaataa accttgtgca gttgtacagt acaaatcatg gaaaggtggt ggaattattg
      901 cagttagcta tgaagcccga gcatttggtg tcactagaaa catgtgggcg gatgatgcta
      961 agaagttatg tccagatctt ctcctggcac aagttcgtga gtcccgtgga aaagctaacc
     1021 tcaccaagta ccgggaagct agtgttgaag tgatggagat aatgtcttat tttgctgtga
     1081 ttgaacgtgc cagcatcgat gaggcttata tagatttgac cagtgctgta caagaaagac
     1141 ttcaaaagtt acaaggtcag cctatctcag cagatttgct gccaagcact tacattgaag
     1201 ggttgccccg aggccctaca gtagaagaga ctgttcagaa agaggctatt cggaaacaag
     1261 gcttgcttca gtggcttgat tctcttcaga gtgatgatcc cacctctcca gacctcaggc
     1321 tgactgtggg agccatgatt gtggaagaaa tgagagcagc catagagagt aagactggct
     1381 tccagtgttc agctggaatc tcacacaata aggtcctggc aaaacttgct tgtggactaa
     1441 ataagcccaa ccgccaaacc ctggtctcac atgggtcagt gccacagctc ttcagccaga
     1501 tgccaatccg taaaatccga agtcttggag gaaagctagg ggcttctgtc attgaagtcc
     1561 ttgggataga atacatgggt gacctgaccc agttcactga atcccagctc cagagtcatt
     1621 ttggagagaa gaatgggtct tggctgtatg ccatgtgtcg aggtattgaa cacgatccag
     1681 ttaaacccag gcagttacct aaaactattg gctgcagcaa gaacttccca gggaaaacag
     1741 ctctggctac tcgggagcag gtacagtggt ggcttttgca gttagctctg gaacttgagg
     1801 agaggctaac caaagatcga aatgataatg accgcgtggc cactcagctg gtcgtcagta
     1861 ttcgttttca gggagacagg cgtctcagca gcctgcggcg gtgctgtgcc ctccctcgct
     1921 atgacgctca caagatgagc caggacgcat ttgctgccat caggaactgt aatacatctg
     1981 gaatccaaac tgagtggtcc cctcctctca caatgctctt cctctgcgct accaagttct
     2041 ctgcggctgc ccctccagct tgcacggata tcactgcgtt cctgagtagc gactcaagtt
     2101 gtcagccaaa ggtgccaatt gccagctctg agactaggac ccaggggtct ggcccagctg
     2161 taccaacctc taaagaggca gccacatctc tagcgtcttt ttttcaaaaa gctgccaaaa
     2221 agcagagaat gaaagagact tcctttgtac ctcttaacac agccacagag aagttatcat
     2281 ccaagccttc acttgtgttc caaagcagcc aaaccacagg aagtcagtct ttctttaagc
     2341 agaagagcct gctgctccag cacacacagc ttagtaactc tgcggctcct gaccctccac
     2401 aggccagccc tgcagcgcaa ccaagctgtc tgccagcaga gtgtgtagac agtggtcctg
     2461 atgatggagc ggtgaagcca gtatcctcta aagcagtgtc tacagaaatg aatgtggctg
     2521 gggacagtcc aaacgtgctc gattctccag cttacaactc tcaggaggtg acccaaaggg
     2581 ccactgagga ccaagtgctc tgtgagaagt gtgactccct ggttccagtg tgggacatgc
     2641 cagaacacac ggactaccat tttgcattgg agttgcagaa gtcttttttg cagccttgta
     2701 cttcaaaacc ccaggctatt cctgccgtgt ctcctcaggg caaaagaaat cccaagagtc
     2761 cctcggcttc cagtagtaag cgccttaggc cccatggcat gcagacgctg gaatcgtttt
     2821 ttaagccact gacacatgga tccgactaca aagaccatga cggtgattat aaagatcatg
     2881 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     2941 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3001 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3061 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3121 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3181 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3241 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3301 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3361 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3421 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3481 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     3541 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3601 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3661 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3721 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3781 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     3841 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3901 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3961 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     4021 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4081 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4141 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4201 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4261 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4621 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4681 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4741 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4801 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4861 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4921 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4981 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5041 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5101 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5161 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5221 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5281 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5341 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5401 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5461 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     5521 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5581 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5641 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5701 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5761 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5821 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5881 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5941 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6001 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7261 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7561 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7801 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7861 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7921 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7981 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     8041 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     8101 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
