LOCUS       V019289                 7324 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019289
VERSION     V019289
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1051..2877
                     /label="ALB(NM_000477)"
                     /note="ALB(NM_000477)"
                     /gene="ALB"
     polyA_signal    2924..3148
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3194..3622
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3636..3965
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4032..4826
                     /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    5000..5121
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5170..5186)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5194..5210
                     /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(5218..5248)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5263..5284
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5572..6157)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6328..7188)
                     /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(7189..7293)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggtaccga gctcggatcc atgaagtggg taacctttat ttcccttctt
     1081 tttctcttta gctcggctta ttccaggggt gtgtttcgtc gagatgcaca caagagtgag
     1141 gttgctcatc ggtttaaaga tttgggagaa gaaaatttca aagccttggt gttgattgcc
     1201 tttgctcagt atcttcagca gtgtccattt gaagatcatg taaaattagt gaatgaagta
     1261 actgaatttg caaaaacatg tgttgctgat gagtcagctg aaaattgtga caaatcactt
     1321 catacccttt ttggagacaa attatgcaca gttgcaactc ttcgtgaaac ctatggtgaa
     1381 atggctgact gctgtgcaaa acaagaacct gagagaaatg aatgcttctt gcaacacaaa
     1441 gatgacaacc caaacctccc ccgattggtg agaccagagg ttgatgtgat gtgcactgct
     1501 tttcatgaca atgaagagac atttttgaaa aaatacttat atgaaattgc cagaagacat
     1561 ccttactttt atgccccgga actccttttc tttgctaaaa ggtataaagc tgcttttaca
     1621 gaatgttgcc aagctgctga taaagctgcc tgcctgttgc caaagctcga tgaacttcgg
     1681 gatgaaggga aggcttcgtc tgccaaacag agactcaagt gtgccagtct ccaaaaattt
     1741 ggagaaagag ctttcaaagc atgggcagta gctcgcctga gccagagatt tcccaaagct
     1801 gagtttgcag aagtttccaa gttagtgaca gatcttacca aagtccacac ggaatgctgc
     1861 catggagatc tgcttgaatg tgctgatgac agggcggacc ttgccaagta tatctgtgaa
     1921 aatcaagatt cgatctccag taaactgaag gaatgctgtg aaaaacctct gttggaaaaa
     1981 tcccactgca ttgccgaagt ggaaaatgat gagatgcctg ctgacttgcc ttcattagct
     2041 gctgattttg ttgaaagtaa ggatgtttgc aaaaactatg ctgaggcaaa ggatgtcttc
     2101 ctgggcatgt ttttgtatga atatgcaaga aggcatcctg attactctgt cgtgctgctg
     2161 ctgagacttg ccaagacata tgaaaccact ctagagaagt gctgtgccgc tgcagatcct
     2221 catgaatgct atgccaaagt gttcgatgaa tttaaacctc ttgtggaaga gcctcagaat
     2281 ttaatcaaac aaaattgtga gctttttgag cagcttggag agtacaaatt ccagaatgcg
     2341 ctattagttc gttacaccaa gaaagtaccc caagtgtcaa ctccaactct tgtagaggtc
     2401 tcaagaaacc taggaaaagt gggcagcaaa tgttgtaaac atcctgaagc aaaaagaatg
     2461 ccctgtgcag aagactatct atccgtggtc ctgaaccagt tatgtgtgtt gcatgagaaa
     2521 acgccagtaa gtgacagagt caccaaatgc tgcacagaat ccttggtgaa caggcgacca
     2581 tgcttttcag ctctggaagt cgatgaaaca tacgttccca aagagtttaa tgctgaaaca
     2641 ttcaccttcc atgcagatat atgcacactt tctgagaagg agagacaaat caagaaacaa
     2701 actgcacttg ttgagctcgt gaaacacaag cccaaggcaa caaaagagca actgaaagct
     2761 gttatggatg atttcgcagc ttttgtagag aagtgctgca aggctgacga taaggagacc
     2821 tgctttgccg aggagggtaa aaaacttgtt gctgcaagtc aagctgcctt aggcttataa
     2881 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2941 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3001 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3061 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3121 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3181 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3241 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3301 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3361 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3421 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3481 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3541 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3601 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3661 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3781 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3841 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3901 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3961 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4021 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4081 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4141 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4201 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4261 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4321 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4381 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4441 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4501 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4561 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4621 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4681 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4741 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4801 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4861 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4921 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4981 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5041 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5101 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5161 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5221 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5281 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5761 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6001 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7321 cgtc
//
