LOCUS       V042843                 8761 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042843
VERSION     V042843
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..3417
                     /label="MCM2(NM_004526)"
                     /note="MCM2(NM_004526)"
                     /gene="MCM2"
     CDS             3424..3513
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3537..4125
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4191..4239
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4441..4869
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4883..5212
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5279..6073
                     /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    6249..6370
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6419..6435)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6443..6459
                     /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(6467..6497)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6512..6533
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6821..7409)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7580..8440)
                     /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(8441..8545)
                     /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 ggaatcatcg
      721 gaatccttca ccatggcatc cagcccggcc cagcgtcggc gaggcaatga tcctctcacc
      781 tccagccctg gccgaagctc ccggcgtact gatgccctca cctccagccc tggccgtgac
      841 cttccaccat ttgaggatga gtccgagggg ctcctaggca cagaggggcc cctggaggaa
      901 gaagaggatg gagaggagct cattggagat ggcatggaaa gggactaccg cgccatccca
      961 gagctggacg cctatgaggc cgagggactg gctctggatg atgaggacgt agaggagctg
     1021 acggccagtc agagggaggc agcagagcgg gccatgcggc agcgtgaccg ggaggctggc
     1081 cggggcctgg gccgcatgcg ccgtgggctc ctgtatgaca gcgatgagga ggacgaggag
     1141 cgccctgccc gcaagcgccg ccaggtggag cgggccacgg aggacggcga ggaggacgag
     1201 gagatgatcg agagcatcga gaacctggag gatctcaaag gccactctgt gcgcgagtgg
     1261 gtgagcatgg cgggcccccg gctggagatc caccaccgct tcaagaactt cctgcgcact
     1321 cacgtcgaca gccacggcca caacgtcttc aaggagcgca tcagcgacat gtgcaaagag
     1381 aaccgtgaga gcctggtggt gaactatgag gacttggcag ccagggagca cgtgctggcc
     1441 tacttcctgc ctgaggcacc ggcggagctg ctgcagatct ttgatgaggc tgccctggag
     1501 gtggtactgg ccatgtaccc caagtacgac cgcatcacca accacatcca tgtccgcatc
     1561 tcccacctgc ctctggtgga ggagctgcgc tcgctgaggc agctgcatct gaaccagctg
     1621 atccgcacca gtggggtggt gaccagctgc actggcgtcc tgccccagct cagcatggtc
     1681 aagtacaact gcaacaagtg caatttcgtc ctgggtcctt tctgccagtc ccagaaccag
     1741 gaggtgaaac caggctcctg tcctgagtgc cagtcggccg gcccctttga ggtcaacatg
     1801 gaggagacca tctatcagaa ctaccagcgt atccgaatcc aggagagtcc aggcaaagtg
     1861 gcggctggcc ggctgccccg ctccaaggac gccattctcc tcgcagatct ggtggacagc
     1921 tgcaagccag gagacgagat agagctgact ggcatctatc acaacaacta tgatggctcc
     1981 ctcaacactg ccaatggctt ccctgtcttt gccactgtca tcctagccaa ccacgtggcc
     2041 aagaaggaca acaaggttgc tgtaggggaa ctgaccgatg aagatgtgaa gatgatcact
     2101 agcctctcca aggatcagca gatcggagag aagatctttg ccagcattgc tccttccatc
     2161 tatggtcatg aagacatcaa gagaggcctg gctctggccc tgttcggagg ggagcccaaa
     2221 aacccaggtg gcaagcacaa ggtacgtggt gatatcaacg tgctcttgtg cggagaccct
     2281 ggcacagcga agtcgcagtt tctcaagtat attgagaaag tgtccagccg agccatcttc
     2341 accactggcc agggggcgtc ggctgtgggc ctcacggcgt atgtccagcg gcaccctgtc
     2401 agcagggagt ggaccttgga ggctggggcc ctggttctgg ctgaccgagg agtgtgtctc
     2461 attgatgaat ttgacaagat gaatgaccag gacagaacca gcatccatga ggccatggag
     2521 caacagagca tctccatctc gaaggctggc atcgtcacct ccctgcaggc tcgctgcacg
     2581 gtcattgctg ccgccaaccc cataggaggg cgctacgacc cctcgctgac tttctctgag
     2641 aacgtggacc tcacagagcc catcatctca cgctttgaca tcctgtgtgt ggtgagggac
     2701 accgtggacc cagtccagga cgagatgctg gcccgcttcg tggtgggcag ccacgtcaga
     2761 caccacccca gcaacaagga ggaggagggg ctggccaatg gcagcgctgc tgagcccgcc
     2821 atgcccaaca cgtatggcgt ggagcccctg ccccaggagg tcctgaagaa gtacatcatc
     2881 tacgccaagg agagggtcca cccgaagctc aaccagatgg accaggacaa ggtggccaag
     2941 atgtacagtg acctgaggaa agaatctatg gcgacaggca gcatccccat tacggtgcgg
     3001 cacatcgagt ccatgatccg catggcggag gcccacgcgc gcatccatct gcgggactat
     3061 gtgatcgaag acgacgtcaa catggccatc cgcgtgatgc tggagagctt catagacaca
     3121 cagaagttca gcgtcatgcg cagcatgcgc aagacttttg cccgctacct ttcattccgg
     3181 cgtgacaaca atgagctgtt gctcttcata ctgaagcagt tagtggcaga gcaggtgaca
     3241 tatcagcgca accgctttgg ggcccagcag gacactattg aggtccctga gaaggacttg
     3301 gtggataagg ctcgtcagat caacatccac aacctctctg cattttatga cagtgagctc
     3361 ttcaggatga acaagttcag ccacgacctg aaaaggaaaa tgatcctgca gcagttcctc
     3421 gaggaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat
     3481 ttagagcaaa agttaatttc cgaggaggac ttatagtgag ggcccgatat ctcgacaatc
     3541 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3601 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3661 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3721 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3781 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3841 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3901 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     3961 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4021 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4081 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     4141 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4201 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4261 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4321 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4381 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     4441 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4501 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4561 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     4621 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4681 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4741 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4801 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4861 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4921 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5041 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5101 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     5161 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     5221 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     5281 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5341 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5401 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5461 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5521 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5581 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5641 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5701 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5761 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5821 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5881 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5941 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6001 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6061 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     6121 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6181 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6241 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6301 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6361 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6661 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6721 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     7021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7261 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8581 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     8641 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8701 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8761 g
//
