LOCUS       V040643                 7071 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040643
VERSION     V040643
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..1751
                     /label="NEIL2(NM_001135746)"
                     /note="NEIL2(NM_001135746)"
                     /gene="NEIL2"
     CDS             1758..1823
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1847..2435
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2501..2549
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2751..3179
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3193..3522
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3589..4383
                     /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    4559..4680
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4729..4745)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4753..4769
                     /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(4777..4807)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4822..4843
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5890..6750)
                     /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(6751..6855)
                     /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 ccaccatgcc agaagggccg ttggtgagga
      781 aatttcacca tttggtctcc ccctttgtgg gtcagcaggt ggtcaagaca gggggcagca
      841 gtaagaagct acagcccgcc agcctgcagt ctctgtggct ccaggacacc caggtccatg
      901 gaaagaaatt attccttaga tttgatctag atgaagaaat ggggccccct ggcagcagcc
      961 caacaccaga gcctccacaa aaagaagtgc agaaggaagg ggctgcggac ccaaagcagg
     1021 tcggggagcc cagcgggcag aagacccttg atggatcctc acggtctgca gagctcgtcc
     1081 cccagggcga ggatgattct gagtatttgg agagagacgc ccctgcagga gatgctggga
     1141 ggtggctgcg tgtcagcttt ggtttgtttg gcagcgtttg ggtgaacgat ttctccagag
     1201 ccaagaaagc caacaagagg ggggactgga gggacccttc cccgaggttg gtcctgcact
     1261 ttggtggtgg tggcttcctg gcattttata attgtcagtt gtcttggagc tcttccccag
     1321 tggtcacacc cacctgtgac atcctgtctg agaagttcca tcgaggacaa gccttagaag
     1381 ctctaggcca ggctcagcct gtctgctata cactgctgga ccagagatac ttctcagggc
     1441 tagggaacat cattaagaat gaagccttgt acagagctgg gatccatccc ctttctctcg
     1501 gttcagtcct gagtgcctcg cgtcgggagg tcctggtgga tcacgtggtg gagttcagta
     1561 cagcctggct gcagggcaag ttccaaggca gaccgcagca cacacaggtc taccagaaag
     1621 aacagtgccc tgctggccac caggtcatga aggaggcgtt tgggcccgaa gatgggttac
     1681 agaggctcac ctggtggtgc ccgcagtgcc agccccagtt gtcagaggag ccagagcagt
     1741 gccagttctc cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     1801 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     1861 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1921 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1981 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2041 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2101 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2161 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2221 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2281 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2341 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2401 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2461 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2521 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2581 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2641 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2701 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2881 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     2941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3121 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3421 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3541 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4441 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4501 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4561 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4621 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4681 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6901 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     6961 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7021 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
