LOCUS       V019092                 8339 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019092
VERSION     V019092
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             671..3010
                     /label="STRN(NM_003162)"
                     /note="STRN(NM_003162)"
                     /gene="STRN"
     CDS             3026..3091
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3115..3703
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3769..3817
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4019..4447
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4461..4790
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4857..5651
                     /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    5827..5948
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5997..6013)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6021..6037
                     /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(6045..6075)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6090..6111
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6399..6987)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7158..8018)
                     /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(8019..8123)
                     /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 taccgccacc atggacgagc aggcgggtcc cggcgtcttc ttcagcaaca accacccggg
      721 cgccggcggt gccaaggggc tcgggcctct ggcggaggct gccgcggccg gcgacggggc
      781 ggctgcggcg ggggcggccc gagcccagta cagtctcccg gggatcctgc acttcctgca
      841 gcacgagtgg gcccgcttcg aggtggagag agcccagtgg gaggtggagc gggcggagct
      901 gcaggcccag attgccttcc tgcagggaga aaggaagggc caagaaaatt tgaagaagga
      961 tcttgtgagg aggatcaaaa tgttggagta tgctcttaaa caggaaagag ccaaatacca
     1021 caagttgaaa tacgggacag aattgaatca gggagatatg aagcctccaa gctatgattc
     1081 tgatgaaggt aatgaaacag aagtgcagcc acaacaaaac agccagttaa tgtggaaaca
     1141 aggtcgacaa ctactcagac agtatctaca ggaggtgggt tatacagata ctattctaga
     1201 tgtgaaatct aaacgagtgc gagctttgtt gggcttttca agtgatgtca cggacaggga
     1261 agatgacaaa aatcaggact cagttgtaaa tggcacagag gctgaagtta aagagacagc
     1321 aatgattgca aaatctgagt taacagattc tgcctccgtg ctggataatt tcaaattcct
     1381 tgaaagtgca gctgcagatt tcagtgatga agatgaagat gatgatgttg atggaagaga
     1441 gaaaagcgtc attgatactt caacaattgt taggaaaaaa gcattgcctg acagcggtga
     1501 agatcgagat acaaaagaag ctctaaagga gtttgacttc ttggttacat cagaggaagg
     1561 agacaatgaa tctagaagtg caggcgatgg aacagactgg gaaaaggaag accagtgtct
     1621 catgcctgaa gcctggaatg tggaccaggg agtaattacc aaactcaagg aacaatacaa
     1681 aaaggagaga aaggggaaaa agggggtgaa gaggcccaat aggtcaaaac tacaagatat
     1741 gcttgctaat ttgagagatg ttgatgaact tccttcattg cagccatctg tgggttcacc
     1801 ttccagaccc agcagctcca ggcttcctga acatgaaatt aatagggcag atgaagtgga
     1861 agcattgaca tttcctcctt cttctggaaa gtcattcatc atgggagcag atgaagccct
     1921 tgaaagtgaa ctgggacttg gagaactagc aggccttacg gtggccaatg aagcagactc
     1981 actaacttat gatatagcaa acaataaaga tgcattgagg aagacatgga accctaagtt
     2041 tacattgaga agtcactttg atggcatccg agcccttgct ttccatccca ttgagcctgt
     2101 tttgataaca gcatcagagg atcacacatt aaaaatgtgg aatttacaga aaacagcccc
     2161 agccaaaaag agcacttctc ttgatgtaga acctatctat acattcagag cccataaagg
     2221 tccagtgctt tgtgtggtaa tgagcagcaa tggtgagcag tgttacagtg gtggtactga
     2281 tggactgatc cagggctgga ataccactaa tcccaacatc gacccctatg attcttatga
     2341 tccttctgtt ttacgaggcc ctctgctagg ccacacggat gcagtctggg gtttggctta
     2401 tagtgcagca catcagcgtt tgttgtcctg ttcagcagat ggcactctgc gtttatggaa
     2461 tacaactgag gttgctccag cactaagtgt atttaatgat actaaagaac tgggaatccc
     2521 tgcctctgtg gatctagtga gcagtgaccc gagccatatg gtagcatcat tcagcaaggg
     2581 atatacaagc atttttaaca tggaaacaca acaacgcatt ctcactttag aatccaatgt
     2641 agatacaaca gccaactctt cctgccaaat aaatagagtc atcagtcatc ctactcttcc
     2701 gatcagcatc actgctcatg aagacaggca catcaaattc tatgataaca atacaggcaa
     2761 actgatccac tcgatggtag cccacctaga agctgttaca agtttagcag ttgatcccaa
     2821 tggcctttac ttgatgtctg gcagtcatga ctgttcaata cgtttatgga atctagaaag
     2881 taagacgtgt atccaagaat tcacagctca tcgaaaaaag tttgaagaat cgattcatga
     2941 tgtagctttc cacccatcca aatgctatat agccagtgct ggagctgacg cactggctaa
     3001 agtctttgta atgcggccgc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3061 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     3121 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3181 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3241 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3301 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3361 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3421 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3481 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     3541 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3601 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3661 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     3721 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3781 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3841 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3901 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3961 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     4021 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4081 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4141 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     4201 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4261 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4321 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4381 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4441 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4621 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4681 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4741 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4801 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4861 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4921 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4981 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5041 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5101 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5161 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5221 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5281 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5341 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5401 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5461 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5521 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5581 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5641 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     5701 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5761 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5821 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5941 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     6001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7441 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7741 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     8161 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     8221 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     8281 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
