LOCUS       V028050                 9117 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028050
VERSION     V028050
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..3797
                     /label="PARP1(NM_001618)"
                     /note="PARP1(NM_001618)"
                     /gene="PARP1"
     CDS             3804..3869
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3893..4481
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4547..4595
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4797..5225
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5239..5568
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5635..6429
                     /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    6605..6726
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6775..6791)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6799..6815
                     /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(6823..6853)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6868..6889
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7177..7765)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7936..8796)
                     /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(8797..8901)
                     /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 ggagtcttcg gataagctct
      781 atcgagtcga gtacgccaag agcgggcgcg cctcttgcaa gaaatgcagc gagagcatcc
      841 ccaaggactc gctccggatg gccatcatgg tgcagtcgcc catgtttgat ggaaaagtcc
      901 cacactggta ccacttctcc tgcttctgga aggtgggcca ctccatccgg caccctgacg
      961 ttgaggtgga tgggttctct gagcttcggt gggatgacca gcagaaagtc aagaagacag
     1021 cggaagctgg aggagtgaca ggcaaaggcc aggatggaat tggtagcaag gcagagaaga
     1081 ctctgggtga ctttgcagca gagtatgcca agtccaacag aagtacgtgc aaggggtgta
     1141 tggagaagat agaaaagggc caggtgcgcc tgtccaagaa gatggtggac ccggagaagc
     1201 cacagctagg catgattgac cgctggtacc atccaggctg ctttgtcaag aacagggagg
     1261 agctgggttt ccggcccgag tacagtgcga gtcagctcaa gggcttcagc ctccttgcta
     1321 cagaggataa agaagccctg aagaagcagc tcccaggagt caagagtgaa ggaaagagaa
     1381 aaggcgatga ggtggatgga gtggatgaag tggcgaagaa gaaatctaaa aaagaaaaag
     1441 acaaggatag taagcttgaa aaagccctaa aggctcagaa cgacctgatc tggaacatca
     1501 aggacgagct aaagaaagtg tgttcaacta atgacctgaa ggagctactc atcttcaaca
     1561 agcagcaagt gccttctggg gagtcggcga tcttggaccg agtagctgat ggcatggtgt
     1621 tcggtgccct ccttccctgc gaggaatgct cgggtcagct ggtcttcaag agcgatgcct
     1681 attactgcac tggggacgtc actgcctgga ccaagtgtat ggtcaagaca cagacaccca
     1741 accggaagga gtgggtaacc ccaaaggaat tccgagaaat ctcttacctc aagaaattga
     1801 aggttaaaaa acaggaccgt atattccccc cagaaaccag cgcctccgtg gcggccacgc
     1861 ctccgccctc cacagcctcg gctcctgctg ctgtgaactc ctctgcttca gcagataagc
     1921 cattatccaa catgaagatc ctgactctcg ggaagctgtc ccggaacaag gatgaagtga
     1981 aggccatgat tgagaaactc ggggggaagt tgacggggac ggccaacaag gcttccctgt
     2041 gcatcagcac caaaaaggag gtggaaaaga tgaataagaa gatggaggaa gtaaaggaag
     2101 ccaacatccg agttgtgtct gaggacttcc tccaggacgt ctccgcctcc accaagagcc
     2161 ttcaggagtt gttcttagcg cacatcttgt ccccttgggg ggcagaggtg aaggcagagc
     2221 ctgttgaagt tgtggcccca agagggaagt caggggctgc gctctccaaa aaaagcaagg
     2281 gccaggtcaa ggaggaaggt atcaacaaat ctgaaaagag aatgaaatta actcttaaag
     2341 gaggagcagc tgtggatcct gattctggac tggaacactc tgcgcatgtc ctggagaaag
     2401 gtgggaaggt cttcagtgcc acccttggcc tggtggacat cgttaaagga accaactcct
     2461 actacaagct gcagcttctg gaggacgaca aggaaaacag gtattggata ttcaggtcct
     2521 ggggccgtgt gggtacggtg atcggtagca acaaactgga acagatgccg tccaaggagg
     2581 atgccattga gcacttcatg aaattatatg aagaaaaaac cgggaacgct tggcactcca
     2641 aaaatttcac gaagtatccc aaaaagttct accccctgga gattgactat ggccaggatg
     2701 aagaggcagt gaagaagctg acagtaaatc ctggcaccaa gtccaagctc cccaagccag
     2761 ttcaggacct catcaagatg atctttgatg tggaaagtat gaagaaagcc atggtggagt
     2821 atgagatcga ccttcagaag atgcccttgg ggaagctgag caaaaggcag atccaggccg
     2881 catactccat cctcagtgag gtccagcagg cggtgtctca gggcagcagc gactctcaga
     2941 tcctggatct ctcaaatcgc ttttacaccc tgatccccca cgactttggg atgaagaagc
     3001 ctccgctcct gaacaatgca gacagtgtgc aggccaaggt ggaaatgctt gacaacctgc
     3061 tggacatcga ggtggcctac agtctgctca ggggagggtc tgatgatagc agcaaggatc
     3121 ccatcgatgt caactatgag aagctcaaaa ctgacattaa ggtggttgac agagattctg
     3181 aagaagccga gatcatcagg aagtatgtta agaacactca tgcaaccaca cacaatgcgt
     3241 atgacttgga agtcatcgat atctttaaga tagagcgtga aggcgaatgc cagcgttaca
     3301 agccctttaa gcagcttcat aaccgaagat tgctgtggca cgggtccagg accaccaact
     3361 ttgctgggat cctgtcccag ggtcttcgga tagccccgcc tgaagcgccc gtgacaggct
     3421 acatgtttgg taaagggatc tatttcgctg acatggtctc caagagtgcc aactactgcc
     3481 atacgtctca gggagaccca ataggcttaa tcctgttggg agaagttgcc cttggaaaca
     3541 tgtatgaact gaagcacgct tcacatatca gcaagttacc caagggcaag cacagtgtca
     3601 aaggtttggg caaaactacc cctgatcctt cagctaacat tagtctggat ggtgtagacg
     3661 ttcctcttgg gaccgggatt tcatctggtg tgaatgacac ctctctacta tataacgagt
     3721 acattgtcta tgatattgct caggtaaatc tgaagtatct gctgaaactg aaattcaatt
     3781 ttaagacctc cctgtgggga tccgactaca aagaccatga cggtgattat aaagatcatg
     3841 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     3901 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3961 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4021 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4081 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4141 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4201 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4261 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4321 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4381 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4441 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4501 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4561 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4621 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4681 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4741 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     4801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4861 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4921 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     4981 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5101 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5161 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5221 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5401 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5461 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5521 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5581 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5641 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5701 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5761 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5821 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5881 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5941 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6001 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6061 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6121 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6181 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6241 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6301 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6361 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6421 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6481 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6601 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6661 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6721 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6781 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6841 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6901 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6961 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7021 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7081 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7141 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7201 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7261 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7321 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7381 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7441 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7501 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7561 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7621 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7681 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7741 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7801 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7861 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7921 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7981 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8041 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8101 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8161 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8221 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8281 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8341 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8401 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8461 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8521 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8581 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8641 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8701 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8761 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8821 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8881 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8941 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     9001 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9061 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
