LOCUS       V039995                 8352 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039995
VERSION     V039995
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             657..3062
                     /label="CDC5L(NM_001253)"
                     /note="CDC5L(NM_001253)"
                     /gene="CDC5L"
     CDS             3078..3095
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    3128..3716
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3782..3830
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4032..4460
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4474..4803
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4870..5664
                     /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    5840..5961
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6010..6026)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6034..6050
                     /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(6058..6088)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6103..6124
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6412..7000)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7171..8031)
                     /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(8032..8136)
                     /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 gctaggatcc gccaccatgc
      661 ctcgaattat gatcaagggg ggcgtatgga ggaataccga ggatgaaatt ctgaaagcag
      721 cggtaatgaa atatgggaaa aatcagtggt ctaggattgc ctcattgctg catagaaaat
      781 cagcaaagca gtgcaaagcc agatggtatg aatggctgga tccaagcatt aagaagacag
      841 aatggtccag agaagaagag gaaaaactct tgcacttggc caagttgatg ccaactcagt
      901 ggaggaccat tgctccaatc attggaagaa cagcggccca gtgcttagaa cactatgaat
      961 ttcttctgga taaagctgcc caaagagaca atgaagagga aacaacagat gatccacgaa
     1021 aacttaaacc tggagaaata gatccaaatc cagaaacaaa accagcgcgg cctgatccaa
     1081 ttgatatgga tgaggatgaa cttgagatgc tttctgaagc cagagcccgc ttggctaata
     1141 ctcagggaaa gaaggccaag aggaaagcaa gagagaaaca attggaagaa gcaagacgtc
     1201 ttgctgccct ccaaaaaaga agagaacttc gagcagctgg catagaaatt cagaagaaaa
     1261 gaaaaaggaa gagaggagtt gattataatg ccgaaatccc atttgaaaaa aagcctgccc
     1321 ttggttttta tgatacttct gaggaaaact accaagctct tgacgcagat ttcaggaaat
     1381 taagacaaca ggatcttgat ggggagctaa gatctgaaaa agaaggaaga gatagaaaaa
     1441 aagacaaaca gcatttgaaa aggaaaaaag aatctgattt accatcagct attcttcaaa
     1501 ctagtggtgt ttctgaattt actaaaaaga gaagcaaact agtacttcct gcccctcaga
     1561 tttcagatgc agaactccag gaagttgtaa aagtaggcca agcgagtgaa attgcacgtc
     1621 aaactgccga ggaatctggc ataacaaatt ctgcttccag tacacttttg tctgagtaca
     1681 atgtcaccaa caacagcgtt gctcttagaa caccacgaac accagcttcc caggacagaa
     1741 ttctgcagga agcccagaac ctcatggccc tcaccaatgt ggacacccca ttgaaaggtg
     1801 gacttaatac cccattgcat gagagtgact tctcaggtgt aactccacag cgacaagttg
     1861 tacagactcc aaacacagtt ctctctactc cattcaggac tccttctaat ggagctgaag
     1921 ggctgactcc ccggagtgga acaactccca aaccagttat taactctact ccgggtagaa
     1981 ctcctcttcg agacaagtta aacattaatc ccgaggatgg aatggcagac tatagtgatc
     2041 cctcttacgt gaagcagatg gaaagagaat cccgagaaca tctccgttta gggttgttgg
     2101 gccttcctgc ccctaagaat gattttgaaa ttgttctacc agaaaatgcc gagaaggagc
     2161 tggaagaacg tgaaatagat gatacttaca ttgaagatgc tgctgatgtg gatgctcgaa
     2221 agcaggccat acgagatgca gagcgtgtaa aggaaatgaa acgaatgcat aaagctgtcc
     2281 agaaagatct gccaagacca tcagaagtaa atgaaactat tctaagaccc ttaaatgtag
     2341 aaccgccttt aacagattta cagaaaagtg aagaactaat caaaaaagaa atgatcacaa
     2401 tgcttcatta tgaccttcta catcaccctt atgaaccatc tggaaataaa aaaggcaaaa
     2461 ctgtagggtt tggtaccaat aattcagagc acattaccta tctggaacat aatccttatg
     2521 aaaagttctc caaagaagag ctgaaaaagg cccaggatgt tttggtgcag gagatggaag
     2581 tggttaaaca aggaatgagc catggagagc tctcaagtga agcttataac caggtgtggg
     2641 aagaatgcta cagtcaagtt ttatatcttc ctgggcagag ccgctacaca cgggccaatc
     2701 tggctagtaa aaaggacaga attgaatcac ttgaaaagag gctcgagata aacaggggtc
     2761 acatgacgac agaagccaag agggctgcaa agatggaaaa gaagatgaaa attttgcttg
     2821 ggggttacca gtctcgtgct atggggctca tgaaacagtt gaatgactta tgggaccaaa
     2881 ttgaacaggc tcacttggag ttacgcactt ttgaagaact caagaaacat gaagattctg
     2941 ctattccccg gaggctagag tgtctaaaag aagacgttca gcgacaacaa gaaagagaaa
     3001 aggaacttca acatagatat gctgatttgc tgctggagaa agagacttta aagtcaaaat
     3061 tcgaattcgc caccatgcac catcaccatc accattagct cgagtctaga gggcccgata
     3121 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3181 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3241 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3301 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3361 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3421 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3481 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3541 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3601 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3661 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3721 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3781 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3841 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3901 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3961 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4021 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4381 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4441 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4741 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4801 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4861 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4921 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4981 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5041 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5101 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5161 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5221 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5281 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5341 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5401 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5461 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5521 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5581 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5641 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5701 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5761 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5821 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5881 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5941 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6061 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6241 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6301 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6601 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6841 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7081 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7141 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7201 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7261 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7321 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7381 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7441 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7501 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7561 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7621 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7681 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7741 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7801 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7861 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7921 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7981 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8041 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8101 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8161 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8221 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8281 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8341 aatctgctta gg
//
