LOCUS       V026272                 7716 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026272
VERSION     V026272
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..2396
                     /label="Scp2(NM_011327)"
                     /note="Scp2(NM_011327)"
                     /gene="Scp2"
     CDS             2403..2468
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2492..3080
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3146..3194
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3396..3824
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3838..4167
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4234..5028
                     /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    5204..5325
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5374..5390)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5398..5414
                     /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(5422..5452)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5467..5488
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5776..6364)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6535..7395)
                     /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(7396..7500)
                     /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 ccaccatgcc ttctgtcgct ttgaaatctc
      781 cgcgcctgcg ccgggtgttc gtggtcggcg ttggcatgac caagttcatg aagcctggag
      841 gtgagaactc aagagactat cctgatatgg caaaggaagc aggccagaag gcattggaag
      901 atgcacagat cccttactcc gcagtggaac aggcatgcgt tggctatgtg tatggtgatt
      961 ccaccagtgg gcagagggct atctatcata gtttgggact gactggcatt cctataatta
     1021 atgtcaacaa taactgttct actggttcga ctgctttgtt tatggcccat caactgattc
     1081 aaggaggctt ggcaaattgt gtgttggctc ttgggtttga gaagatggag aggggatcca
     1141 ttggaacaaa attctcagat cggaccactc caactgataa acatattgaa gtcttgatcg
     1201 ataagtatgg attgtctgca catccgatta ctcctcagat gtttgggtat gctgggaaag
     1261 aacatatgga aaaatatgga acaaaagttg aacactttgc aaaaattgga tggaaaaatc
     1321 ataaacactc agttaataac acgtattccc agttccaaga tgaatacagc ttagaagaag
     1381 taatgaaatc aaaaccagtt ttcgattttc tgactatctt gcaatgctgc cccacctcag
     1441 atggtgccgc tgccgcgatt ctgtccagcg aggagtttgt gcagcagtac ggcctgcagt
     1501 ccaaagcggt ggagattgtg gcccaggaga tgatgactga cttacccagt acgtttgaag
     1561 agaaaagtat tattaaagtg gttggctatg atatgagtaa agaagctgcc aggagatgct
     1621 atgagaagtc cggcctgaca cccaacgatg tcgacgtgat agagcttcac gattgcttct
     1681 ctgtcaatga actcatcact tacgaagcac tggggctctg tccagaagga caaggtggaa
     1741 ccctggtgga cagaggggac aacacttacg gaggaaagtg ggtcatcaac cctagtggag
     1801 gcctcatctc aaagggacac ccactgggcg caacaggtct ggctcagtgc gcggagctct
     1861 gctggcagct gagaggcgaa gccggaaaga ggcaggttcc cggggcaaag gtggctctgc
     1921 agcacaatct aggcctcgga ggagctgtgg ttgtcaccct ctacaggatg ggttttcccg
     1981 aagctgccag ctccttcagg acgcaccagg tttcagctgc tcccaccagc tctgcagggg
     2041 atggattcaa ggcaaacctc gtctttaagg agattgagaa gaagcttgaa gaggaagggg
     2101 aacagttcgt gaagaaaatc ggtggcattt ttgccttcaa agtgaaagat ggccctggag
     2161 gcaaagaagc tacctgggtg gtggatgtga agaatggcaa aggatctgtg cttcccaatt
     2221 cagataagaa ggctgactgc acaatcacca tggccgactc agacttgctg gctttgatga
     2281 ctggaaaaat gaaccctcag tcggccttct ttcaagggaa actgaagatt gctggtaaca
     2341 tgggactggc catgaaacta cagaaccttc agcttcagcc gggcaaagct aagctgggat
     2401 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     2461 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     2521 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2581 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2641 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2701 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2761 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2821 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2881 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2941 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3001 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3061 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3121 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3181 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3241 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3301 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3361 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3421 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3481 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3541 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3601 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3661 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3721 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3781 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3841 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3901 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4021 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4081 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4141 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4201 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4261 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4321 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4381 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4441 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4501 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4561 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4621 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4681 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4741 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4801 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4861 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4921 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4981 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5041 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5101 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5161 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5221 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5281 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5341 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5401 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5461 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5521 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5941 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6181 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6601 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7501 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7561 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7621 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7681 ggcttgaccg acaattgcat gaagaatctg cttagg
//
