LOCUS       V034381                 7804 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034381
VERSION     V034381
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..1722
                     /label="RAD51(NM_002875)"
                     /note="RAD51(NM_002875)"
                     /gene="RAD51"
     CDS             1753..2460
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             2467..2556
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2580..3168
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3234..3282
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3484..3912
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3926..4255
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4322..5116
                     /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    5292..5413
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5462..5478)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5486..5502
                     /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(5510..5540)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5555..5576
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5864..6452)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6623..7483)
                     /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(7484..7588)
                     /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 ccaccatggc aatgcagatg
      721 cagcttgaag caaatgcaga tacttcagtg gaagaagaaa gctttggccc acaacccatt
      781 tcacggttag agcagtgtgg cataaatgcc aacgatgtga agaaattgga agaagctgga
      841 ttccatactg tggaggctgt tgcctatgcg ccaaagaagg agctaataaa tattaaggga
      901 attagtgaag ccaaagctga taaaattctg gctgaggcag ctaaattagt tccaatgggt
      961 ttcaccactg caactgaatt ccaccaaagg cggtcagaga tcatacagat tactactggc
     1021 tccaaagagc ttgacaaact acttcaaggt ggaattgaga ctggatctat cacagaaatg
     1081 tttggagaat tccgaactgg gaagacccag atctgtcata cgctagctgt cacctgccag
     1141 cttcccattg accggggtgg aggtgaagga aaggccatgt acattgacac tgagggtacc
     1201 tttaggccag aacggctgct ggcagtggct gagaggtatg gtctctctgg cagtgatgtc
     1261 ctggataatg tagcatatgc tcgagcgttc aacacagacc accagaccca gctcctttat
     1321 caagcatcag ccatgatggt agaatctagg tatgcactgc ttattgtaga cagtgccacc
     1381 gccctttaca gaacagacta ctcgggtcga ggtgagcttt cagccaggca gatgcacttg
     1441 gccaggtttc tgcggatgct tctgcgactc gctgatgagt ttggtgtagc agtggtaatc
     1501 actaatcagg tggtagctca agtggatgga gcagcgatgt ttgctgctga tcccaaaaaa
     1561 cctattggag gaaatatcat cgcccatgca tcaacaacca gattgtatct gaggaaagga
     1621 agaggggaaa ccagaatctg caaaatctac gactctccct gtcttcctga agctgaagct
     1681 atgttcgcca ttaatgcaga tggagtggga gatgccaaag acggtggagg cggatcagga
     1741 ggcggtggat ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1801 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1861 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     1921 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1981 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2041 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2101 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2161 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2221 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2281 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2341 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2401 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     2461 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa
     2521 gatttagagc aaaagttaat ttccgaggag gacttatagt gagggcccga tatctcgaca
     2581 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2641 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2701 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2761 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2821 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2881 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2941 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3001 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3061 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3121 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     3181 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3241 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3301 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3361 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3421 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     3481 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     3661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3841 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3901 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3961 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4081 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4141 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4201 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4261 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4321 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4381 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4441 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4501 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4561 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4621 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4681 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4741 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4801 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4861 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4921 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4981 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5041 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5101 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     5161 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5221 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5281 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5401 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6301 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7621 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     7681 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7741 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7801 tagg
//
