LOCUS       V037444                 9910 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037444
VERSION     V037444
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             964..5400
                     /label="PXDN(NM_012293)"
                     /note="PXDN(NM_012293)"
                     /gene="PXDN"
     CDS             5407..5472
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5510..5734
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5780..6208
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6222..6551
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6618..7412
                     /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    7586..7707
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7756..7772)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7780..7796
                     /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(7804..7834)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7849..7870
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8158..8743)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8914..9774)
                     /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(9775..9879)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcca agcgctccag gggccccggg cgccgctgcc tgttggcgct cgtgctgttc
     1021 tgcgcctggg ggacgctggc cgtggtggcc cagaagccgg gcgcagggtg tccgagccgc
     1081 tgcctgtgct tccgcaccac cgtgcgctgc atgcatctgc tgctggaggc cgtgcccgcc
     1141 gtggcgccgc agacctccat cctagatctt cgctttaaca gaatcagaga gatccaacct
     1201 ggggcattca ggcggctgag gaacttgaac acattgcttc tcaataataa tcagatcaag
     1261 aggataccta gtggagcatt tgaagacttg gaaaatttaa aatatctcta tctgtacaag
     1321 aatgagatcc agtcaattga caggcaagca tttaagggac ttgcctctct agagcaacta
     1381 tacctgcact ttaatcagat agaaactttg gacccagatt cgttccagca tctcccgaag
     1441 ctcgagaggc tatttttgca taacaaccgg attacacatt tagttccagg gacatttaat
     1501 cacttggaat ctatgaagag attgcgactg gactcaaaca cacttcactg cgactgtgaa
     1561 atcctgtggt tggcggattt gctgaaaacc tacgcggagt cggggaacgc gcaggcagcg
     1621 gccatctgtg aatatcccag acgcatccag ggacgctcag tggcaaccat caccccggaa
     1681 gagctgaact gtgaaaggcc ccggatcacc tccgagcccc aggacgcaga tgtgacctcg
     1741 gggaacaccg tgtacttcac ctgcagagcc gaaggcaacc ccaagcctga gatcatctgg
     1801 ctgcgaaaca ataatgagct gagcatgaag acagattccc gcctaaactt gctggacgat
     1861 gggaccctga tgatccagaa cacacaggag acagaccagg gtatctacca gtgcatggca
     1921 aagaacgtgg ccggagaggt gaagacgcaa gaggtgaccc tcaggtactt cgggtctcca
     1981 gctcgaccca cttttgtaat ccagccacag aatacagagg tgctggttgg ggagagcgtc
     2041 acgctggagt gcagcgccac aggccacccc ccgccgcgga tctcctggac gagaggtgac
     2101 cgcacaccct tgccagttga cccgcgggtg aacatcacgc cttctggcgg gctttacata
     2161 cagaacgtcg tacaggggga cagcggagag tatgcgtgct ctgcgaccaa caacattgac
     2221 agcgtccatg ccaccgcttt catcatcgtc caggctcttc ctcagttcac tgtgacgcct
     2281 caggacagag tcgttattga gggccagacc gtggatttcc agtgtgaagc caagggcaac
     2341 ccgccgcccg tcatcgcctg gaccaaggga gggagccagc tctccgtgga ccggcggcac
     2401 ctggtcctgt catcgggaac acttagaatc tctggtgttg ccctccacga ccagggccag
     2461 tacgaatgcc aggctgtcaa catcatcggc tcccagaagg tcgtggccca cctgactgtg
     2521 cagcccagag tcaccccagt gtttgccagc attcccagcg acacaacagt ggaggtgggc
     2581 gccaatgtgc agctcccgtg cagctcccag ggcgagcccg agccagccat cacctggaac
     2641 aaggatgggg ttcaggtgac agaaagtgga aaatttcaca tcagccctga aggattcttg
     2701 accatcaatg acgttggccc tgcagacgca ggtcgctatg agtgtgtggc ccggaacacc
     2761 attgggtcgg cctcggtgag catggtgctc agtgtgaatg ttcctgacgt cagtcgaaat
     2821 ggagatccgt ttgtagctac ctccatcgtg gaagcgattg cgactgttga cagagctata
     2881 aactcaaccc gaacacattt gtttgacagc cgtcctcgtt ctccaaatga tttgctggcc
     2941 ttgttccggt atccgaggga tccttacaca gttgaacagg cacgggcggg agaaatcttt
     3001 gaacggacat tgcagctcat tcaggagcat gtacagcatg gcttgatggt cgacctcaac
     3061 ggaacaagtt accactacaa cgacctggtg tctccacagt acctgaacct catcgcaaac
     3121 ctgtcgggct gtaccgccca ccggcgcgtg aacaactgct cggacatgtg cttccaccag
     3181 aagtaccgga cgcacgacgg cacctgtaac aacctgcagc accccatgtg gggcgcctcg
     3241 ctgaccgcct tcgagcgcct gctgaaatcc gtgtacgaga atggcttcaa cacccctcgg
     3301 ggcatcaacc cccaccgact gtacaacggg cacgcccttc ccatgccgcg cctggtgtcc
     3361 accaccctga tcgggacgga gaccgtcaca cccgacgagc agttcaccca catgctgatg
     3421 cagtggggcc agttcctgga ccacgacctc gactccacgg tggtggccct gagccaggca
     3481 cgcttctccg acggacagca ctgcagcaac gtgtgcagca acgacccccc ctgcttctct
     3541 gtcatgatcc cccccaatga ctcccgggcc aggagcgggg cccgctgcat gttcttcgtg
     3601 cgctccagcc ctgtgtgcgg cagcggcatg acttcgctgc tcatgaactc cgtgtacccg
     3661 cgggagcaga tcaaccagct cacctcctac atagacgcat ccaacgtgta cgggagcacg
     3721 gagcatgagg cccgcagcat ccgcgacctg gccagccacc gcggcctgct gcggcagggc
     3781 atcgtgcagc ggtccgggaa gccgctgctc cccttcgcca ccgggccgcc cacggagtgc
     3841 atgcgggacg agaacgagag ccccatcccc tgcttcctgg ccggggacca ccgcgccaac
     3901 gagcagctgg gcctgaccag catgcacacg ctgtggttcc gcgagcacaa ccgcattgcc
     3961 acggagctgc tcaagctgaa cccgcactgg gacggcgaca ccatctacta tgagaccagg
     4021 aagatcgtgg gtgcggagat ccagcacatc acctaccagc actggctccc gaagatcctg
     4081 ggggaggtgg gcatgaggac gctgggagag taccacggct acgaccccgg catcaatgct
     4141 ggcatcttca acgccttcgc caccgcggcc ttcaggtttg gccacacgct tgtcaaccca
     4201 ctgctttacc ggctggacga gaacttccag cccattgcac aagatcacct cccccttcac
     4261 aaagctttct tctctccctt ccggattgtg aatgagggcg gcatcgatcc gcttctcagg
     4321 gggctgttcg gggtggcggg gaaaatgcgt gtgccctcgc agctgctgaa cacggagctc
     4381 acggagcggc tgttctccat ggcacacacg gtggctctgg acctggcggc catcaacatc
     4441 cagcggggcc gggaccacgg gatcccaccc taccacgact acagggtcta ctgcaatcta
     4501 tcggcggcac acacgttcga ggacctgaaa aatgagatta aaaaccctga gatccgggag
     4561 aaactgaaaa ggttgtatgg ctcgacactc aacatcgacc tgtttccggc gctcgtggtg
     4621 gaggacctgg tgcctggcag ccggctgggc cccaccctga tgtgtcttct cagcacacag
     4681 ttcaagcgcc tgcgagatgg ggacaggttg tggtatgaga accctggggt gttctccccg
     4741 gcccagctga ctcagatcaa gcagacgtcg ctggccagga tcctatgcga caacgcggac
     4801 aacatcaccc gggtgcagag cgacgtgttc agggtggcgg agttccctca cggctacggc
     4861 agctgtgacg agatccccag ggtagacctc cgggtgtggc aggactgctg tgaagactgt
     4921 aggaccaggg ggcagttcaa tgccttttcc tatcatttcc gaggcagacg gtctcttgag
     4981 ttcagctacc aggaggacaa gccgaccaag aaaacaagac cacggaaaat acccagtgtt
     5041 gggagacagg gggaacatct cagcaacagc acctcagcct tcagcacacg ctcagatgca
     5101 tctgggacaa atgacttcag agagtttgtt ctggaaatgc agaagaccat cacagacctc
     5161 agaacacaga taaagaaact tgaatcacgg ctcagtacca cagagtgcgt ggatgccggg
     5221 ggcgaatctc acgccaacaa caccaagtgg aaaaaagatg catgcaccat ttgtgaatgc
     5281 aaagacgggc aggtcacctg cttcgtggaa gcttgccccc ctgccacctg tgctgtcccc
     5341 gtgaacatcc caggggcctg ctgtccagtc tgcttacaga agagggcgga ggaaaagccc
     5401 ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     5461 gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     5521 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5581 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5641 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5701 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5761 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5821 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6001 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6061 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6121 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6181 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6481 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6541 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     6601 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6781 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     7021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     7081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     7141 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7381 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7441 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7501 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7561 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7621 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7681 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     8701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8881 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8941 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9001 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9061 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9121 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9181 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9241 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9301 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9361 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9421 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9481 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9541 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9601 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9661 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9721 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9781 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9841 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9901 acctgacgtc
//
