LOCUS       V045051                 8292 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045051
VERSION     V045051
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             720..1373
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
                     /label="GST"
     CDS             1410..3002
                     /label="PKM(NM_002654)"
                     /note="PKM(NM_002654)"
                     /gene="PKM"
     misc_feature    3068..3656
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3722..3770
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3972..4400
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4414..4743
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4810..5604
                     /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    5780..5901
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5950..5966)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5974..5990
                     /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(5998..6028)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6043..6064
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6352..6940)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7111..7971)
                     /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(7972..8076)
                     /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 ctagccacca
      721 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
      781 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
      841 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
      901 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
      961 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     1021 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     1081 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     1141 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     1201 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     1261 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     1321 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaaggaggag
     1381 ggggaagcgg cggcggaggg agtaagctta tgtcgaagcc ccatagtgaa gccgggactg
     1441 ccttcattca gacccagcag ctgcacgcag ccatggctga cacattcctg gagcacatgt
     1501 gccgcctgga cattgattca ccacccatca cagcccggaa cactggcatc atctgtacca
     1561 ttggcccagc ttcccgatca gtggagacgt tgaaggagat gattaagtct ggaatgaatg
     1621 tggctcgtct gaacttctct catggaactc atgagtacca tgcggagacc atcaagaatg
     1681 tgcgcacagc cacggaaagc tttgcttctg accccatcct ctaccggccc gttgctgtgg
     1741 ctctagacac taaaggacct gagatccgaa ctgggctcat caagggcagc ggcactgcag
     1801 aggtggagct gaagaaggga gccactctca aaatcacgct ggataacgcc tacatggaaa
     1861 agtgtgacga gaacatcctg tggctggact acaagaacat ctgcaaggtg gtggaagtgg
     1921 gcagcaagat ctacgtggat gatgggctta tttctctcca ggtgaagcag aaaggtgccg
     1981 acttcctggt gacggaggtg gaaaatggtg gctccttggg cagcaagaag ggtgtgaacc
     2041 ttcctggggc tgctgtggac ttgcctgctg tgtcggagaa ggacatccag gatctgaagt
     2101 ttggggtcga gcaggatgtt gatatggtgt ttgcgtcatt catccgcaag gcatctgatg
     2161 tccatgaagt taggaaggtc ctgggagaga agggaaagaa catcaagatt atcagcaaaa
     2221 tcgagaatca tgagggggtt cggaggtttg atgaaatcct ggaggccagt gatgggatca
     2281 tggtggctcg tggtgatcta ggcattgaga ttcctgcaga gaaggtcttc cttgctcaga
     2341 agatgatgat tggacggtgc aaccgagctg ggaagcctgt catctgtgct actcagatgc
     2401 tggagagcat gatcaagaag ccccgcccca ctcgggctga aggcagtgat gtggccaatg
     2461 cagtcctgga tggagccgac tgcatcatgc tgtctggaga aacagccaaa ggggactatc
     2521 ctctggaggc tgtgcgcatg cagcacctga ttgcccgtga ggcagaggct gccatctacc
     2581 acttgcaatt atttgaggaa ctccgccgcc tggcgcccat taccagcgac cccacagaag
     2641 ccaccgccgt gggtgccgtg gaggcctcct tcaagtgctg cagtggggcc ataatcgtcc
     2701 tcaccaagtc tggcaggtct gctcaccagg tggccagata ccgcccacgt gcccccatca
     2761 ttgctgtgac ccggaatccc cagacagctc gtcaggccca cctgtaccgt ggcatcttcc
     2821 ctgtgctgtg caaggaccca gtccaggagg cctgggctga ggacgtggac ctccgggtga
     2881 actttgccat gaatgttggc aaggcccgag gcttcttcaa gaagggagat gtggtcattg
     2941 tgctgaccgg atggcgccct ggctccggct tcaccaacac catgcgtgtt gttcctgtgc
     3001 cgtgagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgata
     3061 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3121 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3181 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3241 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3301 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3361 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3421 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3481 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3541 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3601 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3661 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3721 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3781 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3841 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3901 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3961 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4021 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4081 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4141 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4201 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4261 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4321 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4381 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4501 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4561 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4621 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4681 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4741 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4801 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4861 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4921 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4981 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5041 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5101 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5161 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5221 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5281 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5341 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5401 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5461 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5521 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5581 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5641 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5761 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5881 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5941 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6001 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6061 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6121 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6181 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6241 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6301 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6541 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6601 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6661 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6721 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6781 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6841 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7021 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7081 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7141 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7201 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7261 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7321 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7381 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7441 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7501 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7561 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7621 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7681 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7741 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7801 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7861 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7921 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7981 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8041 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8101 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8161 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8221 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8281 aatctgctta gg
//
