LOCUS       V019021                 8393 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019021
VERSION     V019021
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..3073
                     /label="STRN3(NM_001083893)"
                     /note="STRN3(NM_001083893)"
                     /gene="STRN3"
     CDS             3080..3145
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3169..3757
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3823..3871
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4073..4501
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4515..4844
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4911..5705
                     /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    5881..6002
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6051..6067)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6075..6091
                     /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(6099..6129)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6144..6165
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6453..7041)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7212..8072)
                     /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(8073..8177)
                     /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 ccatggacga gcttgccgga ggcggtggtg gcggcccggg
      721 gatggcggcc cctccccggc agcagcaggg acctgggggg aacctgggcc tttcgcccgg
      781 ggggaacgga gcggcgggcg gcgggggtcc tccggcctcc gagggagcgg gtcccgcggc
      841 aggccccgag ctgtcccggc cgcagcagta cactatcccg gggatactgc actacatcca
      901 gcacgagtgg gctcggttcg agatggagcg ggcgcactgg gaggtggaac gggccgaact
      961 gcaggcccgg attgcatttc tacaaggcga aagaaaaggt caagagaacc tgaagaagga
     1021 cttagtaaga agaataaaga tgttagagta tgcattaaaa caagaaaggg caaaatatca
     1081 caaattaaaa tatggcacgg aactgaacca aggtgacttg aaaatgccaa cctttgagtc
     1141 agaagaaacc aaagacacag aggctcccac agcacctcag aatagccagt taacgtggaa
     1201 gcaaggcaga cagcttttaa gacagtatct tcaggaagta ggttatacag atacaatatt
     1261 agatgtacgg tctcagcggg taaggtcatt acttggacta tctaattcag aaccaaatgg
     1321 atcagtagaa acaaagaatt tagaacagat cctgaatgga ggtgaatctc ctaagcaaaa
     1381 gggacaagaa ataaaaaggt cctctggtga tgttcttgag acgttcaatt tcttagaaaa
     1441 tgccgatgac agtgatgaag atgaggaaaa tgacatgatc gaaggcatcc cagaaggaaa
     1501 agacaaacat cggatgaata aacataaaat aggtaatgaa ggtttagctg ctgacctaac
     1561 tgacgatcct gatactgagg aagcactgaa agaatttgat tttttagtga ctgctgaaga
     1621 tggtgaagga gctggagaag cacggagttc gggggatggc acagaatggg ataaagatga
     1681 cctctcccca actgctgagg tttgggatgt agaccaggga ctaataagta aactgaagga
     1741 acagtacaag aaggaacgaa aggggaagaa aggggtgaag agggccaaca ggacaaaact
     1801 ctacgacatg atagctgatc tgggagatga tgagctgccc cacatccctt caggaatcat
     1861 taatcagtct aggtcagcct ctactagaat gactgatcat gaaggtgcaa gagcagagga
     1921 agctgaacca ataacgtttc catctggagg aggcaagtca tttattatgg gttctgatga
     1981 tgttttgtta agtgtactgg gccttggaga ccttgcagac ttgacggtaa caaatgatgc
     2041 agactatagt tatgatttgc ctgctaataa agatgccttt cgaaagacat ggaatcccaa
     2101 gtatacacta cgtagccatt ttgatggagt acgggcatta gcttttcatc ctgtagaacc
     2161 tgtgctggtt actgcttctg aggaccatac cctgaaactt tggaacctgc aaaaaacagt
     2221 tcctgccaaa aagagtgcct ctttagatgt agagcctatc tacacattta gggcccacat
     2281 cggccctgtt ctgtcattag ctattagttc taatggagaa cagtgtttta gtggtggtat
     2341 tgatgcaacc atccagtggt ggaatatgcc gagtcccagt gtagatccat atgatacata
     2401 tgagccaaat gttctagctg gcactttagt tggtcataca gatgcagttt ggggtcttgc
     2461 ttatagtggc ataaaaaatc aattactgtc ttgttcagca gatggcactg ttaggttatg
     2521 gaatccacaa gaaaaattgc catgtatttg cacttacaat ggagataaaa agcatggaat
     2581 acctacatca gttgacttta taggctgtga tccagctcat atggtaacct ctttcaacac
     2641 tggtagtgca gtaatttatg atttagaaac atcacagtca ttggtgatac tttcatcaca
     2701 ggtagattct ggtttacaat ctaataatca tatcaacaga gtagtaagtc atcccacact
     2761 tcctgttaca ataactgctc atgaagatag acacatcaaa ttttttgaca ataaaacggg
     2821 taaaatgatc cattctatgg tagctcactt ggatgctgtt acaagtctag cagtagatcc
     2881 taatggaatc tatttgatgt ctggaagcca tgactgttcc atcagattat ggaatttaga
     2941 cagcaagaca tgtgtgcaag aaataacagc tcacagaaag aaattggatg aatcaattta
     3001 tgatgttgct ttccactcgt caaaagcata tatagctagt gcaggagctg atgctcttgc
     3061 caaagtattt gtactcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     3121 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     3181 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3241 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3301 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3361 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3421 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3481 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3541 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3601 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3661 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3721 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     3781 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3841 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3901 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3961 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4021 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     4081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4201 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4261 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4381 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4441 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4501 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4561 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4621 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4681 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4741 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4801 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4861 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4921 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4981 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5041 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5101 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5161 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5221 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5281 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5341 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5401 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5461 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5521 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5581 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5641 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5701 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     5761 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5821 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5881 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5941 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6001 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6061 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7321 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8041 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8101 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8161 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8221 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     8281 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8341 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
