LOCUS       V020331                 7014 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020331
VERSION     V020331
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             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1000..2535
                     /label="POLR1G(NM_001297590)"
                     /note="POLR1G(NM_001297590)"
                     /gene="POLR1G"
     promoter        complement(2566..2584)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2610..2834
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2880..3308
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3322..3652
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3719..4513
                     /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    4687..4808
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4857..4873)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4881..4897
                     /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(4905..4935)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4950..4971
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5259..5847)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6018..6878)
                     /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(6879..6983)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggagacccgc caccatgtac
      901 ccatacgacg tcccagacta cgctacttat ccttatgatg tacctgatta tgctagctat
      961 ccttatgatg tcccagacta cgcatcagct ggaagcttca tggaggagcc ccaggccggc
     1021 ggtgaggatg ctgctcggtt ctcttgtccc cccaacttta ccgcgaagcc cccagcctca
     1081 gagtcccctc gtttctcctt ggaggcgctg acgggtccag atacggagct gtggcttatt
     1141 caggcccctg cagactttgc cccagaatgc ttcaatgggc ggcatgtgcc tctctctggc
     1201 tcccagatcg tcaagggcaa attggcaggc aagcggcacc gctatcgagt cctcagcagc
     1261 tgtccccaag ctggagaagc gaccctgctg gccccctcaa cggaggcagg aggtggactc
     1321 acctgtgcct cagcccccca gggcacccta aggatccttg agggtcccca gcaatccctg
     1381 tcagggagcc ctctgcagcc catcccagca agtcccccac cacagatccc tcctggcctg
     1441 aggcctcggt tctgtgcctt tgggggcaac ccaccagtca cagggcctag gtcagccttg
     1501 gcccccaacc tgctcacctc agggaagaag aaaaaggaga tgcaggtgac agaggcccca
     1561 gtcactcagg aggcagtgaa tgggcacggg gccctggagg tggacatggc tttggggtcg
     1621 ccagaaatgg atgtgcggaa gaagaagaag aaaaaaaatc agcagctgaa agaaccagag
     1681 gcagcagggc ctgtggggac agagcccaca gtggagacac tggagcctct gggagtgctg
     1741 ttcccgtcca ccaccaagaa gaggaagaag cccaaaggga aagaaacctt cgagccagaa
     1801 gacaagacag tgaagcagga acagattaac actgagcctc tagaagacac agtcctgtcc
     1861 ccgaccaaaa agagaaagag gcaaaagggg acggaaggga tggagccaga ggagggggtg
     1921 acagttgagt ctcagccaca ggtgaaggtg gagccactgg aggaagccat ccctctgccc
     1981 cctacgaaga agaggaaaaa agaaaaggga cagatggcaa tgatggagcc agggacggag
     2041 gcgatggagc cagtggagcc ggagatgaag cctctggagt ccccaggggg gaccatggcg
     2101 cctcaacagc cagaaggagc gaagcctcag gcccaggcag ctctggcagc tcccaaaaag
     2161 aagacgaaga aagaaaaaca gcaagatgcc acagtggagc cagagacaga ggtggtgggg
     2221 cctgagctgc cggatgacct tgagcctcag gcagctccca catccaccaa gaagaagaag
     2281 aagaagaaag agagaggtca cacagtgact gagccaattc agccactaga gcctgaactg
     2341 ccaggggagg gacagcctga agccagggca actccgggat ccaccaagaa gaggaagaag
     2401 cagagtcagg aaagccggat gccagagaca gtgccccaag aggagatgcc agggccgcca
     2461 ctgaattcag agtctgggga ggaggctccc acaggccggg acaagaagcg gaagcagcag
     2521 cagcagcagc ctgtgtagct cgagcatgca tctagagggc cctattctat agtgtcacct
     2581 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2641 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2701 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2761 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2821 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2881 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2941 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3001 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3061 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3241 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3301 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3361 ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3481 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3541 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3601 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3661 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3721 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3781 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3841 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3901 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3961 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4021 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4081 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4141 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4201 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4261 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4321 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4381 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4441 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4501 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4561 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4681 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4741 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4801 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4861 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4921 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4981 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5041 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5101 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5161 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5221 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5281 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5341 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5401 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5461 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5521 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5581 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5641 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5701 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5761 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5821 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5881 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
