LOCUS       V037651                 8311 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037651
VERSION     V037651
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..2967
                     /label="NBN(NM_002485)"
                     /note="NBN(NM_002485)"
                     /gene="NBN"
     CDS             2974..3063
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3087..3675
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3741..3789
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3991..4419
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4433..4762
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4829..5623
                     /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    5799..5920
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5969..5985)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5993..6009
                     /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(6017..6047)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6062..6083
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6371..6959)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7130..7990)
                     /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(7991..8095)
                     /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 ccaccatgtg gaaactgctg
      721 cccgccgcgg gcccggcagg aggagaacca tacagacttt tgactggcgt tgagtacgtt
      781 gttggaagga aaaactgtgc cattctgatt gaaaatgatc agtcgatcag ccgaaatcat
      841 gctgtgttaa ctgctaactt ttctgtaacc aacctgagtc aaacagatga aatccctgta
      901 ttgacattaa aagataattc taagtatggt acctttgtta atgaggaaaa aatgcagaat
      961 ggcttttccc gaactttgaa gtcgggggat ggtattactt ttggagtgtt tggaagtaaa
     1021 ttcagaatag agtatgagcc tttggttgca tgctcttctt gtttagatgt ctctgggaaa
     1081 actgctttaa atcaagctat attgcaactt ggaggattta ctgtaaacaa ttggacagaa
     1141 gaatgcactc accttgtcat ggtatcagtg aaagttacca ttaaaacaat atgtgcactc
     1201 atttgtggac gtccaattgt aaagccagaa tattttactg aattcctgaa agcagttgag
     1261 tccaagaagc agcctccaca aattgaaagt ttttacccac ctcttgatga accatctatt
     1321 ggaagtaaaa atgttgatct gtcaggacgg caggaaagaa aacaaatctt caaagggaaa
     1381 acatttatat ttttgaatgc caaacagcat aagaaattga gttccgcagt tgtctttgga
     1441 ggtggggaag ctaggttgat aacagaagag aatgaagaag aacataattt ctttttggct
     1501 ccgggaacgt gtgttgttga tacaggaata acaaactcac agaccttaat tcctgactgt
     1561 cagaagaaat ggattcagtc aataatggat atgctccaaa ggcaaggtct tagacctatt
     1621 cctgaagcag aaattggatt ggcggtgatt ttcatgacta caaagaatta ctgtgatcct
     1681 cagggccatc ccagtacagg attaaagaca acaactccag gaccaagcct ttcacaaggc
     1741 gtgtcagttg atgaaaaact aatgccaagc gccccagtga acactacaac atacgtagct
     1801 gacacagaat cagagcaagc agatacatgg gatttgagtg aaaggccaaa agaaatcaaa
     1861 gtctccaaaa tggaacaaaa attcagaatg ctttcacaag atgcacccac tgtaaaggag
     1921 tcctgcaaaa caagctctaa taataatagt atggtatcaa atactttggc taagatgaga
     1981 atcccaaact atcagctttc accaactaaa ttgccaagta taaataaaag taaagatagg
     2041 gcttctcagc agcagcagac caactccatc agaaactact ttcagccgtc taccaaaaaa
     2101 agggaaaggg atgaagaaaa tcaagaaatg tcttcatgca aatcagcaag aatagaaacg
     2161 tcttgttctc ttttagaaca aacacaacct gctacaccct cattgtggaa aaataaggag
     2221 cagcatctat ctgagaatga gcctgtggac acaaactcag acaataactt atttacagat
     2281 acagatttaa aatctattgt gaaaaattct gccagtaaat ctcatgctgc agaaaagcta
     2341 agatcaaata aaaaaaggga aatggatgat gtggccatag aagatgaagt attggaacag
     2401 ttattcaagg acacaaaacc agagttagaa attgatgtga aagttcaaaa acaggaggaa
     2461 gatgtcaatg ttagaaaaag gccaaggatg gatatagaaa caaatgacac tttcagtgat
     2521 gaagcagtac cagaaagtag caaaatatct caagaaaatg aaattgggaa gaaacgtgaa
     2581 ctcaaggaag actcactatg gtcagctaaa gaaatatcta acaatgacaa acttcaggat
     2641 gatagtgaga tgcttccaaa aaagctgtta ttgactgaat ttagatcact ggtgattaaa
     2701 aactctactt ccagaaatcc atctggcata aatgatgatt atggtcaact aaaaaatttc
     2761 aagaaattca aaaaggtcac atatcctgga gcaggaaaac ttccacacat cattggagga
     2821 tcagatctaa tagctcatca tgctcgaaag aatacagaac tagaagagtg gctaaggcag
     2881 gaaatggagg tacaaaatca acatgcaaaa gaagagtctc ttgctgatga tctttttaga
     2941 tacaatcctt atttaaaaag gagaagactc gaggaacaaa aactaatatc agaggaagat
     3001 ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc cgaggaggac
     3061 ttatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     3121 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3181 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3241 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3301 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3361 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3421 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3481 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3541 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3601 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3661 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     3721 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     3781 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3841 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3901 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3961 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4021 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4081 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4141 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4201 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4261 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4321 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     4381 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4441 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4501 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4561 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4621 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4681 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4741 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4801 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4861 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4921 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4981 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5041 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5101 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5161 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5221 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5281 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5341 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5401 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5461 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5521 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5581 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5641 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5701 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5761 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5821 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5881 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5941 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6181 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6241 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6301 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6541 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6781 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8101 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     8161 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     8221 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     8281 gaccgacaat tgcatgaaga atctgcttag g
//
