LOCUS       V020095                 7484 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020095
VERSION     V020095
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             683..2164
                     /label="ITGBL1(NM_004791)"
                     /note="ITGBL1(NM_004791)"
                     /gene="ITGBL1"
     CDS             2171..2236
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2260..2848
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2914..2962
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3164..3592
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3606..3935
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4002..4796
                     /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    4972..5093
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5142..5158)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5166..5182
                     /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(5190..5220)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5235..5256
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5544..6132)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6303..7163)
                     /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(7164..7268)
                     /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 ggatccgcca ccatgcgtcc cccaggcttc aggaacttct tgctgctggc
      721 gtcctccctt ctctttgctg ggttgtcagc tgttcctcaa agcttctcgc catctctgag
      781 gagctggccg ggcgccgcct gcaggctgtc ccgggccgag tcggagcgac gctgccgcgc
      841 acctgggcag cccccggggg ccgcgctgtg ccacggccgg ggccgctgcg actgcggcgt
      901 ctgcatctgc cacgtgactg agccgggcat gttcttcggg cccctgtgtg agtgccatga
      961 gtgggtgtgc gagacctacg acgggagcac ctgtgcaggc catggtaagt gtgactgtgg
     1021 caagtgcaag tgtgaccagg gatggtatgg ggatgcttgc cagtacccaa ctaactgtga
     1081 cttgacaaag aagaaaagta accaaatgtg caagaattca caagacatca tctgctctaa
     1141 tgcaggtaca tgtcactgtg gcaggtgtaa gtgtgataat tcagatggaa gtggacttgt
     1201 gtatggtaaa ttttgtgagt gtgacgatag agaatgcata gacgatgaaa cagaagaaat
     1261 atgtggaggc catgggaagt gttactgtgg aaactgctac tgcaaggctg gttggcatgg
     1321 agataaatgt gaattccagt gcgatatcac cccctgggaa agcaagcgaa gatgcacgtc
     1381 tccagatggc aaaatctgca gtaacagagg gacttgtgta tgtggtgaat gtacctgtca
     1441 cgatgttgat ccgactgggg actggggaga tattcatggg gacacctgtg aatgtgatga
     1501 gagggactgt agagctgtct atgaccgata ttctgatgac ttctgttcag gtcatggaca
     1561 gtgtaattgc ggaagatgtg actgcaaagc aggctggtat gggaagaagt gtgagcaccc
     1621 acagtcctgc acgctgtcag ctgaggagag catcaggaag tgccagggaa gctcggatct
     1681 gccttgctct gggaggggta aatgtgaatg tggcaaatgc acctgctatc ctccaggaga
     1741 tcgccgggtg tatggcaaga cttgtgagtg tgatgatcgc cgctgtgaag acctcgatgg
     1801 tgtggtctgt ggaggccacg gcacatgttc ctgtggtcgc tgtgtttgtg agagaggatg
     1861 gtttggaaag ctctgccaac atccgcggaa gtgtaacatg acggaagaac aaagcaagaa
     1921 tctgtgtgaa tcagcagatg gcatattgtg ctcggggaag ggttcttgtc attgtgggaa
     1981 gtgcatttgt tctgctgaag agtggtatat ttctggggag ttctgtgact gtgatgacag
     2041 agactgcgac aaacatgatg gtctcatttg tacagggaat ggaatatgta gctgtggaaa
     2101 ctgtgaatgc tgggatggat ggaatggaaa tgcatgtgaa atctggcttg gctcagaata
     2161 tcctctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     2221 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     2281 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2341 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2401 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2461 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2521 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2581 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2641 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2701 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2761 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2821 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     2881 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2941 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3001 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3061 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3121 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3181 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3241 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3301 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3361 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3421 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3481 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3541 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3841 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3901 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3961 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4801 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4861 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5101 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5161 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5221 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5281 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5341 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5401 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5461 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5521 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5581 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5641 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5701 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5761 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5881 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5941 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6001 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6061 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6121 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6181 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6241 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6301 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6361 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6421 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6481 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6721 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6961 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7141 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7201 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7261 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7321 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7381 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7441 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
