LOCUS       V027430                 7017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027430
VERSION     V027430
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1697
                     /label="Sfrp1(NM_001276712)"
                     /note="Sfrp1(NM_001276712)"
                     /gene="Sfrp1"
     CDS             1704..1769
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1793..2381
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2447..2495
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2697..3125
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3139..3468
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3535..4329
                     /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    4505..4626
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4675..4691)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4699..4715
                     /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(4723..4753)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4768..4789
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5077..5665)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5836..6696)
                     /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(6697..6801)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg cgtcgggcgc aacgcgcggg
      781 gtcgcggcgg ggccgcctcc ggcgtgctgc tggcgttggc cgccgcgctg ctggccgcgg
      841 gttcggccag cgagtacgac tacgtgagct tccagtcgga tatcggctcc tatcagagcg
      901 ggcgcttcta caccaagccc ccgcagtgcg tggacatccc ggtggacctg cggctgtgcc
      961 acaacgtggg ctacaagaag atggtgctgc ccaacctgct ggagcacgag accatggcag
     1021 aggtgaagca gcaggccagc agctgggtgc cgctgctcaa caagaactgc cacatgggca
     1081 cccaggtctt cctctgctcg ctcttcgcgc ccgtctgtct ggaccggcct atctacccgt
     1141 gtcgctggct ctgcgaggct gttcgcgact cgtgcgagcc ggtcatgcag ttcttcggct
     1201 tctactggcc cgagatgctc aaatgtgaca agttccccga gggcgacgtc tgcatcgcca
     1261 tgaccccgcc caatgccacc gaagcctcga agccccaagg tacaacagtg tgtcctccgt
     1321 gtgacaacga gttgaaatcg gaggccatca tcgaacatct ctgtgcaagc gagtttgcgc
     1381 tgagaatgaa aatcaaagaa gtaaaaaagg aaaacggtga caagaagatt gtccccaaga
     1441 agaagaaacc cttgaagctg gggcccatca agaagaagga gctgaagcgg ctcgtgcttt
     1501 tcctaaagaa tggcgccgac tgtccctgcc accagctgga caacctcagc cacaacttcc
     1561 tcatcatggg gcgcaaggtg aagagccagt acttgctcac agccattcac aagtgggaca
     1621 agaaaaacaa ggagttcaaa aacttcatga agaaaatgaa aaaccacgag tgtcccacct
     1681 tccagtctgt ttttaaggga tccgactaca aagaccatga cggtgattat aaagatcatg
     1741 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1801 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1861 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1921 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1981 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2041 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2101 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2161 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2221 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2281 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2341 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2401 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2461 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2521 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2581 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2641 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2821 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2881 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3001 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3061 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3121 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3481 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3541 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3601 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3661 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3721 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3781 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3841 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3901 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3961 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4021 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4081 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4141 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4201 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4261 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4321 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6841 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6901 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6961 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
