LOCUS       V018235                 8649 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018235
VERSION     V018235
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..3329
                     /label="Secisbp2(NM_029279)"
                     /note="Secisbp2(NM_029279)"
                     /gene="Secisbp2"
     CDS             3336..3401
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3425..4013
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4079..4127
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4329..4757
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4771..5100
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5167..5961
                     /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    6137..6258
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6307..6323)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6331..6347
                     /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(6355..6385)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6400..6421
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6709..7297)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7468..8328)
                     /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(8329..8433)
                     /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 ccaccatggc gtcggagcgg ccgcgggagc
      781 cggacggcga ggatagcatc aagttgtcag ctgatgtcaa accattcgtc cctaagtttg
      841 ctgggctcaa cgtggcgtgg tcagagtcct cagagacacg tgtcttccca ggctgtgcgg
      901 ccacctacta tccatttgta caggaaccac cagcagctga acagaaaatg tatcccgaag
      961 acatggcttt cggagccccc acctttccag cacagtacgt gtcttctgag atagcgctgc
     1021 atccttttgc ctatcccact tacaccctag agtccgcaca gagtgtttgc tcagtgccaa
     1081 ccctgcagta cgactacagc caagcacggt gtcacccagg ctttcggaca gcaaagcccc
     1141 ggcatgagca cgtgtgccct ccacctcagg aagcaaaagg tgtatttaag aaaaaaccct
     1201 ctgatgagag aagagcatgt gaagagcaaa agtcaagcag cagaagggct gacaatgcgg
     1261 tgccctgtga ggcgagacca gccagggggt ccagtcacct gtcctctcga actgagagca
     1321 gtttgaaatc tgatggttac cacaagcgac ccgaccgcaa gtccagaatc cttgcgaaga
     1381 gtgcatctac ctctaaacct gaatttgagt ttagtaggtt agactttcct gaactgcaga
     1441 gtccaaagaa cagtaacatg ccagagacac agaagccgcc caggtggggg cctcttggcc
     1501 ctgctgccag taacatgcct ctcctaggag acgtcggcaa gcccgtcgca gatatggtag
     1561 agggcaaaat ggtgaagagc gatcacactg atggagctgt gaccagtaat gccactacca
     1621 gttccccttc atgtacccaa gagttgtctt ggacaccaat gggttatatt gttcggcaga
     1681 cagtgtcttc agattcagca gcagccactg aaaatgtgac ttccatgata aacctaaaga
     1741 agactacttc atcagctgat gctaaaaatg ttagtgtgac atctgaggct ttatcttcaa
     1801 atccttccta caacagagaa aagcgtgttt atcctgctcc aaaggccaaa gcatcacaag
     1861 gaggtgaact tgaacaaaac gaaagctcca aaaagaataa gaaaaagaaa gagaagtcta
     1921 aaccgagtta tgaagtcctg acggttcagg agccgccaag gattgaagat gcagaggaat
     1981 tccccaacct gtcagttgcg tcggaaagaa gacacagagg gcaatcaccg aagcttcaca
     2041 gtaaacagca gacgcagaat gaatttaaaa caagtgggaa gaagagccag gtcccagtgc
     2101 agctggacct ggggggcatg ctggccgcgc tggagaagca gcagcagcag cagcacgcct
     2161 cgcacgccaa gccatcctcc agacccgtcg tgttctcagt tggagcagtg ccagtcctgt
     2221 ccaaggatgc ctcctccagt gagaggggac gccgctccag tcagatgaag accccacaca
     2281 accccctgga ctccagtgcc cccctgatga agaaggggaa gcagagggag atacctaagg
     2341 ccaagaagcc cacctcactg aagaagataa ttttgaaaga acggcaagag aggatgcagc
     2401 agcgactcca agaaagtgct gtgagcctga cggtggccag tgatgactca caggatgtgg
     2461 agagtggcgc cagtaaccaa acccccagtc aggacaaccc cacaggtcca gagaagacag
     2521 aagaatcagt gtcttctaca cctgtggttg agggtgagtc agaggagcca gctggcacag
     2581 agttccagag ggacccagag gcttgccagc ctgcccctga cagtgccacc ttccccaaga
     2641 tccacagccg gaggttccgg gactactgca gccagatgct tagtaaagaa gtagatgctt
     2701 gtgtcacggg tctgctcaag gagctggtgc gcttccaaga ccgcatgtac cagaaggatc
     2761 ccgtcaaggc caagacaaaa cggcggctcg tgctggggct gagggaggtc ctgaaacacc
     2821 tgaagctcag gaagctgaag tgcatcatca tctctcccaa ctgtgagaag acccagtcta
     2881 aaggtgggct ggacgacacg ctgcacacca tcatcgactg cgcctgcgag cagaacatcc
     2941 ccttcgtgtt tgcactcaat cgcaaggctc tggggcggag cctgaataaa gctgttcctg
     3001 tcagcattgt agggatcttc agctacgatg gggcccagga ccagttccac aagatggttg
     3061 agctgaccat ggcagcccgt caggcataca agaccatgtt ggagacaatg cggcaggaac
     3121 aggcaggaga acctggacct cagtcccctc ccagcccacc catgcaagac cccatcccat
     3181 ccacggaaga aggcaccctc ccttccactg gagaagagcc acactacatt gagatttgga
     3241 aaaagcacct ggaagcgtac agtcagcgtg ccctggagct ggaagactca ctggaggcgt
     3301 caacctctca gatgatgaac ttgaatttag gatccgacta caaagaccat gacggtgatt
     3361 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     3421 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3481 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3541 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3601 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3661 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3721 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3781 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3841 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3901 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3961 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4021 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4081 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4141 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4201 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4261 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4321 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4381 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4441 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     4501 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4561 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4621 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4681 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4741 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4801 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4921 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4981 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5041 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5101 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5161 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5221 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5281 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5341 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5401 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5461 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5521 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5581 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5641 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5701 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5761 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5821 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5881 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5941 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6001 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6061 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6121 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6241 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6301 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6361 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6421 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6481 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6541 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6601 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6661 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6721 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6901 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7141 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7321 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7381 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7441 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7501 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7561 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7621 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7681 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7741 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7801 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7861 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7921 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7981 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8041 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8101 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8161 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8221 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8281 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8341 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8461 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     8521 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     8581 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8641 ctgcttagg
//
