LOCUS       V031757                 9160 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031757
VERSION     V031757
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..3819
                     /label="PITRM1(NM_001242307)"
                     /note="PITRM1(NM_001242307)"
                     /gene="PITRM1"
     CDS             3826..3852
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3856..3882
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3886..3912
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3936..4524
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4590..4638
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4840..5268
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5282..5611
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5678..6472
                     /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    6648..6769
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6818..6834)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6842..6858
                     /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(6866..6896)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6911..6932
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7220..7808)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7979..8839)
                     /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(8840..8944)
                     /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 ccaccatgtg gcgctgcggc
      721 gggcggcagg gcctgtgtgt gctgaggcgg ctgagcggcg gacatgcaca ccacagagcg
      781 tggcgatgga acagtaaccg ggcttgtgag agggctctgc agtataaact aggagacaag
      841 atccatggat tcaccgtaaa ccaggtgaca tctgttcccg agctgttcct gactgcagtg
      901 aagctcaccc atgatgacac aggagccagg tatttacacc tggccagaga agacacgaat
      961 aatctgttca gcgtgcagtt ccgtactact cccatggaca gtactggtgt tcctcacatt
     1021 cttgagcata ccgtcctttg tgggtctcag aaatatccgt gcagagaccc tttcttcaaa
     1081 atgttgaacc ggtccctctc cacgttcatg aacgccttca cagctagtga ttatactctg
     1141 tatccatttt ccacacaaaa tcccaaggac tttcagaatc tcctctcggt gtatttggat
     1201 gccacctttt tcccatgttt acgcgagctg gatttctggc aggaaggatg gcggctggaa
     1261 catgagaatc cgagcgaccc ccagacgccc ttggtcttta aaggagtcgt ctttaatgag
     1321 atgaagggag cgtttacaga caatgagagg atattctccc agcaccttca gaacagactt
     1381 cttcctgacc acacgtactc agtggtctcc gggggtgacc cactgtgcat cccggagctt
     1441 acatgggagc agcttaagca gtttcatgcc actcactatc acccaagcaa tgctaggttc
     1501 ttcacgtacg gtaattttcc attagaacag catctgaaac aaattcacga ggaagcactg
     1561 agcaaattcc agaaaattga accaagcacc gtggtgccag ctcagacacc ctgggacaag
     1621 cctagggaat tccagataac atgtggcccg gattcatttg ctacagatcc ctctaaacaa
     1681 acaaccatca gcgttagctt cctcttaccg gacatcaccg acacatttga agccttcaca
     1741 ttaagtcttc tgtcttcact cttgacttct gggcccaatt ctccctttta caaagccttg
     1801 attgaatctg gccttggcac agacttttct cctgatgttg gatataatgg ctacacgagg
     1861 gaggcctact ttagtgtcgg cctccaaggg attgcggaga aagacattga gaccgtcaga
     1921 agcctcatag acagaacgat tgatgaagta gttgagaaag gatttgaaga tgatcgaatt
     1981 gaggctttac ttcataaaat tgaaatacag atgaaacatc agtctaccag ctttgggctg
     2041 atgctgacat catacatagc ttcttgctgg aaccatgatg gggaccctgt ggagctcttg
     2101 aagttgggaa atcagttagc taaattcaga cagtgcctgc aggaaaatcc aaaatttttg
     2161 caagaaaaag taaaacagta ttttaagaat aaccagcata agctgacttt atcgatgagg
     2221 ccagatgaca agtatcacga gaagcaggca caggtggaag ccacgaagct caagcagaag
     2281 gtcgaggctc tgtcccccgg agacaggcag cagatctacg agaaaggtct agaattacgg
     2341 agtcaacaaa gcaaacctca agatgcctct tgtctgccag cgttgaaagt ttccgatatt
     2401 gaacccacca tacctgtcac agagttggac gtggtcctga cagctggaga tatccctgtt
     2461 cagtactgcg cccagcccac caatggcatg gtgtatttcc gggccttctc cagcctgaac
     2521 acactccccg aggagctgag gccctatgtg cccctcttct gcagcgtcct caccaagctg
     2581 ggctgcggcc ttcttgacta ccgggagcag gctcagcaga tagaattgaa gaccggaggg
     2641 atgagtgctt ctccccacgt gctccccgac gactcacaca tggacaccta cgagcaggta
     2701 ggtgtgcttt tctcctctct ctgcctggat cgaaacctgc cagacatgat gcagctatgg
     2761 agtgaaatat ttaacaaccc gtgctttgaa gaagaggagc acttcaaggt gctggtgaag
     2821 atgaccgccc aggagctcgc caatggaatt cctgactctg ggcacctgta cgcatccatc
     2881 agggcaggcc ggaccctcac gcccgcaggg gacctgcagg agaccttcag cgggatggat
     2941 caggtgcggc tgatgaagag gattgcagaa atgacagata tcaaacccat cctgaggaag
     3001 ctcccgcgta tcaagaaaca cttgttaaat ggtgataata tgaggtgttc agtgaatgcg
     3061 actcctcagc agatgcctca gacagaaaaa gcggtcgaag acttccttag aagcatcggt
     3121 cggagtaaaa aggaacggag gcctgtgcgc ccacacacgg tcgagaaacc tgtgcccagc
     3181 agctctggtg gagatgccca cgttccccat ggctcccagg tcattaggaa gctggtcatg
     3241 gaacccacct tcaagccctg gcagatgaag actcacttcc tgatgccctt cccggtgaat
     3301 tacgtgggtg aatgcatccg aactgtcccc tacacggacc cagatcatgc cagtcttaaa
     3361 atccttgcac gtttgatgac tgccaaattc ttgcatacag aaattcgaga aaaaggcggt
     3421 gcttatggtg gaggcgcaaa actcagccac aatgggattt tcacccttta ctcttacagg
     3481 gacccaaata caatagagac gctccagtct tttgggaagg ctgtcgactg ggctaagtct
     3541 ggaaaattca cacagcaaga catcgacgaa gccaaacttt ctgtcttctc aaccgtagat
     3601 gctcctgtcg ctccttcaga caaaggaatg gaccacttct tgtacggcct ctcggatgag
     3661 atgaagcagg cccacagaga gcagctcttt gctgtcagcc acgacaagct cctggccgtg
     3721 agcgatagat acctcggcac tgggaagagc acacacggcc tggccatcct cggacccgag
     3781 aacccgaaaa ttgccaagga cccatcctgg atcatccaac tcgagtaccc atacgacgta
     3841 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     3901 ccagactacg catagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt
     3961 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4021 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4081 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4141 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4201 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4261 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4321 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4381 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4441 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4501 tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga
     4561 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4621 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4681 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4741 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4801 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4981 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     5221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     5281 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5341 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5401 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5461 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5521 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5581 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5641 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5701 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5761 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5821 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5881 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5941 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6001 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6061 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6121 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6181 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     6241 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6301 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6361 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6421 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6481 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6541 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6601 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6781 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6901 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6961 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7021 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7081 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7141 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7201 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7381 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     7441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7621 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8941 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     9001 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     9061 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     9121 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
