LOCUS       V020446                 8027 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020446
VERSION     V020446
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             683..2707
                     /label="Pros1(NM_031086)"
                     /note="Pros1(NM_031086)"
                     /gene="Pros1"
     CDS             2714..2779
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2803..3391
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3457..3505
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3707..4135
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4149..4478
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4545..5339
                     /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    5515..5636
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5685..5701)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5709..5725
                     /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(5733..5763)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5778..5799
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6087..6675)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6846..7706)
                     /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(7707..7811)
                     /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 ggatccgcca ccatgagggt cctgagcgtg cgctttcggg tgctactggc
      721 ctgcctcgcc ctggtgctcc ctgtctcaga gacaaacttc ttgtcaaaag aacgagcttc
      781 acaagtcctg gtgaggaagc gccgtgcaaa taccttgctc gaagaaacta aaaagggcaa
      841 ccttgaaaga gagtgcattg aagaactctg caataaggag gaagccaggg aggtctttga
      901 aaataatccc gaaacggatt atttttatcc aaaatattta ggttgtctgg gcgcattccg
      961 agttggcgca ttcagtgctg cgaggcagtc agctaatgct taccctgacc tcaggagctg
     1021 tgtcaatgcc atcccagacc aatgtgaccc tatgccatgc aatgaagatg ggtatctgag
     1081 ctgcaaagac ggccaaggtg cttttacttg catctgtaaa ccaggctggc aaggagacaa
     1141 gtgtcagttt gatataaatg aatgcaaaga tccctcaaat atcaatgggg gctgtagcca
     1201 gacttgtgac aatactcctg gaagttacca ctgctcctgt aaaattggct ttgctatgct
     1261 cacaaataag aaagactgca aagatgtgga tgagtgctct ctaaagccaa gtgtctgtgg
     1321 cacagctgtg tgcaagaaca ttccaggaga cttcgaatgt gaatgtccca atggctacag
     1381 atatgatccc tcatcaaaat cttgtaaaga tgtcgatgaa tgctctgaaa acacctgtgc
     1441 ccaactgtgt gtcaattacc ccggaggcta ctcttgttac tgtgatggga agaaaggatt
     1501 caaacttgcc caagatcaga ggagttgtga gggtattcca gtgtgccttt ccttggacct
     1561 tgacaaaaat tatgaattat tatatttggc ggaacagttc gcaggagttg tcttatatct
     1621 gaaatttcga ttgccagata tcaccaggtt ttcagctgaa tttgattttc ggacatatga
     1681 ttcagagggc atcatactgt atgcagaatc tctcgatcac tcaaattggc tcctgattgc
     1741 acttcgtgaa gggaagattg aagttcagtt taagaatgag ttttcaaccc aaatcacaac
     1801 tggaggcaat gttattaaca atggtatatg gaatatggtg tctgtggaag aattagacga
     1861 cagtgttagc attaaaatag ctaaggaggc cgtgatgaat attaataaac ttgggagtct
     1921 ttttaaacct accgatggat ttctggacac caaaatatac tttgcaggat tacctcggaa
     1981 ggtggaaagt gcactcatta agccgattaa tcctcgtctg gatggatgta tacgaggctg
     2041 gaacttgatg aaacaaggag ctttgggtgc aaaggaaata gttgaaggaa aacaaaataa
     2101 acattgcttc ctcactgtgg agaagggctc ctactaccct ggttcaggaa ttgctcagtt
     2161 cagcatagac tacaataatg taactaatgc agagggttgg caaataaatg tgaccttgaa
     2221 tattcgcccg tccactggca ctggagtcat gcttgcttta gtttctgggg acacagtgcc
     2281 ctttgccttg tccttggtgg attctggctc tggaacttct caggacattc tggtatttgt
     2341 tgaaaattca gtagcagctc acttagaagc cataactctg tgctcggaac agccatccca
     2401 gctgaaatgt aacattaaca gaaatggact ggaactgtgg accccagtta gaaaagacgt
     2461 catttactct aaagatctcc aaaggcaact cgccatcttg gacaaaacaa tgaaaggaac
     2521 cgtggccact tacctgggtg gcgttccaga tatttccttc agtgccacac cagtgaatgc
     2581 tttttacagc ggctgcatgg aagtgaacat caacggggta cagttggatc tggatgaagc
     2641 catttccaaa cataatgaca ttagagctca ctcctgtccg tcagtgagga aaatccagaa
     2701 gaacttcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2761 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     2821 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2881 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2941 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3001 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3061 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3121 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3181 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     3241 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3301 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3361 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     3421 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3481 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3541 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3601 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3661 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     3721 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3781 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3841 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     3901 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3961 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4021 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4081 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4141 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4321 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4381 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4441 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4501 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4561 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4621 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4681 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4741 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4801 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4861 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4921 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4981 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5041 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5101 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5161 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5221 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5281 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5341 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5401 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5461 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5521 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5581 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5641 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5701 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5761 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5821 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5881 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5941 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6001 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7741 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7801 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7861 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7921 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7981 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
