LOCUS       V018752                 7871 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018752
VERSION     V018752
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             923..1576
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
                     /label="GST"
     CDS             1586..1603
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
     CDS             1616..3421
                     /label="CKAP4(NM_006825)"
                     /note="CKAP4(NM_006825)"
                     /gene="CKAP4"
     polyA_signal    3468..3692
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3738..4166
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4180..4509
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4576..5370
                     /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    5544..5665
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5714..5730)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5738..5754
                     /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(5762..5792)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5807..5828
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6116..6704)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6875..7735)
                     /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(7736..7840)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgtcccc tatactaggt tattggaaaa ttaagggcct
      961 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
     1021 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
     1081 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
     1141 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
     1201 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
     1261 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
     1321 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
     1381 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
     1441 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
     1501 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
     1561 cgaccatcct ccaaaatcgg atctgctggt gccgcgcggc agcggatccg aattcatgcc
     1621 ctcggccaaa caaaggggct ccaagggcgg ccacggcgcc gcgagcccct cggagaaggg
     1681 tgcccacccg tcgggcggcg cggatgacgt ggcgaagaag ccgccgccgg cgccgcagca
     1741 gccgccgccg ccgcccgcgc cgcacccgca gcagcacccg cagcagcacc cgcagaacca
     1801 ggcgcacggc aagggcggcc accgcggcgg cggcggcggc ggcggcaagt cctcctcctc
     1861 ctcctccgcc tccgccgccg ctgccgccgc cgccgcctcg tcctcggcgt cctgctcgcg
     1921 caggctcggc agggcgctca actttctctt ctacctcgcc ctggtggcgg cggccgcttt
     1981 ctcgggctgg tgcgtccacc acgtcctgga ggaggtccag caggtccggc gcagccacca
     2041 ggacttctcc cggcagaggg aggagctggg ccagggcttg cagggcgtcg agcagaaggt
     2101 gcagtctttg caagccacat ttggaacttt tgagtccatc ttgagaagct cccaacataa
     2161 acaagacctc acagagaaag ctgtgaagca aggggagagt gaggtcagcc ggatcagcga
     2221 agtgctgcag aaactccaga atgagattct caaagacctc tcggatggga tccatgtggt
     2281 gaaggacgcc cgggagcggg acttcacgtc cctggagaac acggtggagg agcggctgac
     2341 ggagctcacc aaatccatca acgacaacat cgccatcttc acagaagtcc agaagaggag
     2401 ccagaaggag atcaatgaca tgaaggcaaa ggttgcctcc ctggaagaat ctgaggggaa
     2461 caagcaggat ttgaaagcct taaaggaagc tgtgaaggag atacagacct cagccaagtc
     2521 cagagagtgg gacatggagg ccctgagaag tacccttcag actatggagt ctgacatcta
     2581 caccgaggtc cgcgagctgg tgagcctcaa gcaggagcag caggctttca aggaggcggc
     2641 cgacacggag cggctcgccc tgcaggccct cacggagaag cttctcaggt ctgaggagtc
     2701 cgtctcccgc ctcccggagg agatccggag actggaggaa gagctccgcc agctgaagtc
     2761 cgattcccac gggccgaagg aggacggagg cttcagacac tcggaagcct ttgaggcact
     2821 ccagcaaaag agtcagggac tggactccag gctccagcac gtggaggatg gggtgctctc
     2881 catgcaggtg gcttctgcgc gccagaccga gagcctggag tccctcctgt ccaagagcca
     2941 ggagcacgag cagcgcctgg ccgccctgca ggggcgcctg gaaggcctcg ggtcctcaga
     3001 ggcagaccag gatggcctgg ccagcacggt gaggagcctg ggcgagaccc agctggtgct
     3061 ctacggtgac gtggaggagc tgaagaggag tgtgggcgag ctccccagca ccgtggaatc
     3121 actccagaag gtgcaggagc aggtgcacac gctgctcagt caggaccaag cccaggccgc
     3181 ccgtctgcct cctcaggact tcctggacag actttcttct ctagacaacc tgaaagcctc
     3241 agtcagccaa gtggaggcgg acttgaaaat gctcaggact gctgtggaca gtttggttgc
     3301 atactcggtc aaaatagaaa ccaacgagaa caatctggaa tcagccaagg gtttactaga
     3361 tgacctgagg aatgatctgg ataggttgtt tgtgaaagtg gagaagattc acgaaaaggt
     3421 ctaactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3481 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3541 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3601 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3661 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3721 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4081 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4141 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4261 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4321 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4381 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4441 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4501 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4561 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4621 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4681 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4741 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4801 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4861 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4921 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4981 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5041 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5101 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5161 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5221 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5281 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5341 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5401 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5461 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5521 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5581 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5641 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6781 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7861 cacctgacgt c
//
