LOCUS       V046927                 7261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046927
VERSION     V046927
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             712..1944
                     /label="STAC2(NM_198993)"
                     /note="STAC2(NM_198993)"
                     /gene="STAC2"
     CDS             1951..2016
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2037..2625
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2691..2739
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2941..3369
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3383..3712
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3779..4573
                     /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    4749..4870
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4919..4935)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4943..4959
                     /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(4967..4997)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5012..5033
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5321..5909)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6080..6940)
                     /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(6941..7045)
                     /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 gcgttttcta gaaaacttaa
      661 gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattcgccac catgaccgag
      721 atgagcgaga aggagaacga accggatgac gcggccaccc acagcccccc agggaccgtc
      781 tccgccctcc aggaaaccaa gctccagcga ttcaagcgct ccctctccct caagaccatc
      841 ctccgaagta agagcttgga gaacttcttc cttcgctcgg gctctgagct caagtgcccc
      901 accgaggtgc tgctgacgcc cccaacccca ctgccccctc cctccccacc acccacagcc
      961 tcggacaggg gcctggctac cccatccccc tccccatgcc cagtcccacg ccccctggca
     1021 gcgctcaaac cagtgaggct gcacagcttc caggaacatg tcttcaagcg agctagccct
     1081 tgtgagctgt gccaccagct catcgtagga aactccaaac agggcttgcg atgtaagatg
     1141 tgcaaagtca gcgtccacct ctggtgctct gaggagatct cccaccagca atgcccaggc
     1201 aagacgtcca cctccttccg ccgcaacttc agttcccctc tcctggtgca tgagccgcca
     1261 ccagtctgtg ccacaagcaa agagtcccca cccactgggg acagtgggaa ggtggaccct
     1321 gtctacgaga ccctgcgcta tggcacctcc ctggcactga tgaaccgctc cagtttcagc
     1381 agcacctctg agtccccgac aaggagcctg agtgagcggg atgagctgac cgaggatggg
     1441 gaaggcagca tccgcagctc tgaggagggg cctggtgaca gtgcatctcc agtattcaca
     1501 gccccagcag agagtgaagg gccaggacca gaggagaaga gtcctggaca gcagctcccc
     1561 aaagccaccc tgcggaagga tgtggggccc atgtactcct acgttgcact ctacaagttt
     1621 ctgccccagg agaacaatga tctggctctg cagcctggag atcggatcat gctggtggat
     1681 gactctaacg aggactggtg gaagggcaag atcggcgacc gggttggctt cttcccagct
     1741 aattttgtgc aacgggtgag gccaggcgag aatgtttggc gctgctgcca acccttctcc
     1801 gggaacaagg aacagggtta catgagcctc aaggagaacc agatctgcgt gggcgtgggc
     1861 agaagcaagg atgctgacgg cttcatccgc gtcagcagtg gcaagaagcg gggcctggtg
     1921 ccagtcgacg ccctgactga gatcctcgag gactacaaag accatgacgg tgattataaa
     1981 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccat ctcgacaatc
     2041 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2101 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2161 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2221 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2281 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2341 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2401 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2461 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2521 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2581 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     2641 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     2701 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     2761 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     2821 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     2881 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3061 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3121 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3241 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3301 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3361 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3721 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3781 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3841 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3901 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3961 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4021 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4081 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4141 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4201 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4261 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4321 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4381 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4441 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4501 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4561 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     4621 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4681 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4741 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4801 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4861 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4921 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4981 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5041 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5101 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5761 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5821 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7081 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     7141 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7201 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7261 g
//
