LOCUS       V041989                 8344 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041989
VERSION     V041989
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             706..3000
                     /label="SEC23A(NM_006364)"
                     /note="SEC23A(NM_006364)"
                     /gene="SEC23A"
     CDS             3007..3096
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3120..3708
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3774..3822
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4024..4452
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4466..4795
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4862..5656
                     /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    5832..5953
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6002..6018)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6026..6042
                     /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(6050..6080)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6095..6116
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6404..6992)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7163..8023)
                     /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(8024..8128)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgac aacctatttg
      721 gaattcattc aacaaaatga agaacgagat ggagtccgat ttagttggaa tgtttggcca
      781 tcaagtcgac tggaagctac aagaatggtt gttcctgtgg cagccctgtt tacaccactg
      841 aaagagagac ctgacttacc acctattcaa tatgaacctg ttctgtgtag taggaccact
      901 tgccgtgcag ttttgaatcc tttatgtcaa gtggattatc gagcaaaact ttgggcttgc
      961 aacttttgtt accaaaggaa tcagtttcca cctagttatg ctggtatatc tgaactgaat
     1021 cagcctgctg aacttttacc tcagttttct agcattgaat atgtagttct gcgtggtcct
     1081 cagatgcctt tgatattcct ctatgtggtt gatacttgca tggaagatga agatttacaa
     1141 gccctgaaag aatccatgca gatgtcatta agtcttttac cacctacagc tttggttgga
     1201 cttattactt ttgggagaat ggttcaggtt catgaacttg gatgtgaagg catttcaaaa
     1261 agctatgtct tcagaggaac aaaagatttg tctgccaaac aactgcagga aatgctgggg
     1321 ctctctaaag taccacttac tcaagcaaca cgtggtcctc aggtacagca gccacctcct
     1381 tccaacagat tcttacaacc agtacagaaa atagacatga atctcacaga tcttctggga
     1441 gaactccagc gagacccttg gcctgtacca cagggaaaga gacctttgcg ttcctctggg
     1501 gtggcacttt ccatagctgt aggactgctg gagtgtactt ttcccaacac tggtgctcgt
     1561 atcatgatgt tcattggtgg tcctgctact caggggcctg gaatggtggt tggagatgag
     1621 ttgaagacac ctataagatc gtggcatgac attgacaaag acaatgccaa atatgttaaa
     1681 aagggaacta agcattttga agcattggct aatcgagctg ctacaactgg ccatgttatt
     1741 gatatctatg cgtgtgcatt agatcagaca ggtctcctgg agatgaaatg ctgtcccaac
     1801 cttactggag gatacatggt aatgggtgat tctttcaata cttccttatt caaacaaact
     1861 tttcaaagag tctttaccaa agacatgcat ggacagttta aaatgggctt tggtggtacg
     1921 ctagaaataa agacctcaag ggaaataaag atttcaggag ctattggacc ctgtgtgtca
     1981 ctcaattcta aaggaccctg tgtgtctgaa aatgagatag gaacaggtgg cacatgtcag
     2041 tggaagatat gtggacttag tcccactaca accttagcca tatattttga ggttgtcaat
     2101 cagcataatg ctccaattcc tcaaggaggg cgtggtgcaa tccagtttgt gactcagtat
     2161 cagcattcaa gtgggcagag acgcatccga gtgaccacca ttgctaggaa ctgggcagat
     2221 gctcaaactc aaatccaaaa cattgctgca tcttttgacc aggaggcagc tgccattctt
     2281 atggcccggc tagcaatata tagagcagaa acagaagaag gtccagatgt gcttaggtgg
     2341 ctggacagac agctcattcg actgtgtcag aaatttggag aatatcataa agatgaccca
     2401 agttccttca gattttcaga aactttctcc ctttatccac agtttatgtt tcatttaaga
     2461 agatcttctt tcctgcaagt ttttaacaat agtcctgatg agagttcata ttatcgtcac
     2521 cattttatgc gtcaagatct gacccagtct ctaattatga ttcagcctat cctgtatgcg
     2581 tattctttta gtggaccacc agagccggtt cttcttgata gcagtagcat tcttgcagat
     2641 cgtattcttc tcatggacac attcttccag attttgattt atcatggtga gaccatagca
     2701 cagtggcgga agtcaggata ccaggatatg cctgagtatg aaaatttccg ccaccttctg
     2761 caagccccag tggatgatgc acaggaaatt cttcactcca gatttccaat gccaagatac
     2821 attgacactg aacatggagg cagccaggcc cgtttcctcc tttcaaaagt caacccttca
     2881 cagactcata ataatatgta tgcctggggg caggagtctg gagcacctat tcttacagat
     2941 gatgttagtt tacaagtgtt tatggatcac ttgaagaaac ttgctgtgtc cagtgctgct
     3001 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa
     3061 gatttagagc aaaagttaat ttccgaggag gacttatagt gagggcccga tatctcgaca
     3121 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3181 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3241 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3301 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3361 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3421 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3481 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3541 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3601 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3661 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     3721 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3781 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3841 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3901 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3961 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     4021 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4081 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4141 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     4201 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4261 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4321 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4381 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4441 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4501 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4561 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4621 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4681 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4741 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4801 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4861 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4921 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4981 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5041 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5101 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5161 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5221 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5281 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5341 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5401 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5461 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5521 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5581 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5641 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     5701 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5761 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5821 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5881 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5941 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6001 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6061 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6121 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6181 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6241 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6301 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6361 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6421 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6481 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6541 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6601 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6661 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6721 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6781 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6841 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6901 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7081 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7141 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7201 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7261 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7321 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7381 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7441 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7501 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7561 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7621 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7681 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7741 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7801 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7861 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7921 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7981 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8041 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8161 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     8221 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     8281 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     8341 tagg
//
