LOCUS       V047078                 7527 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047078
VERSION     V047078
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..2207
                     /label="Irx5(NM_018826)"
                     /note="Irx5(NM_018826)"
                     /gene="Irx5"
     CDS             2214..2279
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2303..2891
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2957..3005
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3207..3635
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3649..3978
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4045..4839
                     /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    5015..5136
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5185..5201)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5209..5225
                     /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(5233..5263)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5278..5299
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5587..6175)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6346..7206)
                     /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(7207..7311)
                     /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 ccaccatgtc ctacccgcag ggctacttgt
      781 accagccgtc cgcctcgctg gcgctctact cgtgccccgc atacagcacc agcgtcattt
      841 cggggccccg cacggatgag ctcggccgct cttcttcggg ctccgcgttc tcgccctacg
      901 ctggctccac tgccttcacg gctccctcgc ccggctacaa ctcgcacctc cagtacggcg
      961 ccgaccccgc ggctgcggcc gccgctgcct tctcttacgt gggctctccc tacgaccaca
     1021 ctcccggtat ggcaggttcc ttgggatatc acccttatgc agcacctctg ggctcctatc
     1081 cttatgggga ccccgcgtac cggaagaatg ccactcggga cgctacagcc actctcaagg
     1141 cctggctcaa cgagcaccgc aagaacccgt accccaccaa gggagagaag atcatgctgg
     1201 ccattatcac caagatgacc ctcacccagg tgtccacctg gtttgccaac gcgcgccggc
     1261 gcctcaagaa agagaacaag atgacgtgga cgccacggaa ccgcagcgag gacgaggagg
     1321 aggaggagaa catcgatctg gagaagaacg acgaggacga gccacagaag cccgaggaca
     1381 agggcgacct cgagggcccc gaatcaggag gagcagagca gaaggcgact gcgggctgcg
     1441 aacggctgca ggggccgctc agccccgccg gcaaggagac cgaaggcagc ctcagcgact
     1501 cggattttaa ggagtcgtcc tccgaaggcc gccacgatga gctgcccagg cccccgcgcg
     1561 cgggcgagtc ctccccggct gggcccgcaa ccgctcgtct ggcggaagac gcgggtcctc
     1621 actaccccgc gagtgtgccg gctcccggcc cgcacccatc agccggagag ctgccccccg
     1681 gctcgggcgg atcctcggtc atccactcgc caccgccacc tccgccaccg cctccggccg
     1741 tgctagccaa gcccaaactg tggtctctgg ccgagatcgc cacatcctcg gacaaggtca
     1801 aggacggggg cggagggagc gagggctctc catgcccacc gtgcccaggg cccatgggcg
     1861 ggcaaacgct tggaggcagc cgcgcgtctc ctgccccagc gcccgcacgc tcaccctccg
     1921 cgcagtgtcc ctttccgggc gggacggtgc tgtcccggcc tctttattac acggcgccct
     1981 tctatccggg ctacacgaac tatggctcct tcggacatct tcacggccac ccagggccag
     2041 ggccaagccc tacagcgggt cctggctctc atttcaatgg attaaaccag acggtgttga
     2101 atcgtgcgga cgttttggct aaagacccga aaatgttgcg gagccagtct cagctagacc
     2161 tgtgcaaaga ctccccctat gagttgaaga aaggtatgtc cgacattgga tccgactaca
     2221 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2281 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2821 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2881 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     2941 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3001 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3061 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3121 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3181 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3361 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3421 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3541 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3601 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3901 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3961 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4021 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4081 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4141 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4201 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4261 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4321 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4381 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4441 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4501 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4561 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4621 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4681 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4741 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4801 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4861 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4921 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4981 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5161 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5761 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6001 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6301 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6361 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6421 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6481 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6541 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6601 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6661 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6721 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6781 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6841 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6901 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6961 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7021 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7081 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7141 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7201 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7261 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7321 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7381 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7441 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7501 gacaattgca tgaagaatct gcttagg
//
