LOCUS       V020014                 8396 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020014
VERSION     V020014
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..3076
                     /label="ITGB1(NM_002211)"
                     /note="ITGB1(NM_002211)"
                     /gene="ITGB1"
     CDS             3083..3148
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3172..3760
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3826..3874
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4076..4504
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4518..4847
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4914..5708
                     /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    5884..6005
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6054..6070)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6078..6094
                     /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(6102..6132)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6147..6168
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6456..7044)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7215..8075)
                     /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(8076..8180)
                     /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 ccatgaattt acaaccaatt ttctggattg gactgatcag
      721 ttcagtttgc tgtgtgtttg ctcaaacaga tgaaaataga tgtttaaaag caaatgccaa
      781 atcatgtgga gaatgtatac aagcagggcc aaattgtggg tggtgcacaa attcaacatt
      841 tttacaggaa ggaatgccta cttctgcacg atgtgatgat ttagaagcct taaaaaagaa
      901 gggttgccct ccagatgaca tagaaaatcc cagaggctcc aaagatataa agaaaaataa
      961 aaatgtaacc aaccgtagca aaggaacagc agagaagctc aagccagagg atattactca
     1021 gatccaacca cagcagttgg ttttgcgatt aagatcaggg gagccacaga catttacatt
     1081 aaaattcaag agagctgaag actatcccat tgacctctac taccttatgg acctgtctta
     1141 ctcaatgaaa gacgatttgg agaatgtaaa aagtcttgga acagatctga tgaatgaaat
     1201 gaggaggatt acttcggact tcagaattgg atttggctca tttgtggaaa agactgtgat
     1261 gccttacatt agcacaacac cagctaagct caggaaccct tgcacaagtg aacagaactg
     1321 caccagccca tttagctaca aaaatgtgct cagtcttact aataaaggag aagtatttaa
     1381 tgaacttgtt ggaaaacagc gcatatctgg aaatttggat tctccagaag gtggtttcga
     1441 tgccatcatg caagttgcag tttgtggatc actgattggc tggaggaatg ttacacggct
     1501 gctggtgttt tccacagatg ccgggtttca ctttgctgga gatgggaaac ttggtggcat
     1561 tgttttacca aatgatggac aatgtcacct ggaaaataat atgtacacaa tgagccatta
     1621 ttatgattat ccttctattg ctcaccttgt ccagaaactg agtgaaaata atattcagac
     1681 aatttttgca gttactgaag aatttcagcc tgtttacaag gagctgaaaa acttgatccc
     1741 taagtcagca gtaggaacat tatctgcaaa ttctagcaat gtaattcagt tgatcattga
     1801 tgcatacaat tccctttcct cagaagtcat tttggaaaac ggcaaattgt cagaaggcgt
     1861 aacaataagt tacaaatctt actgcaagaa cggggtgaat ggaacagggg aaaatggaag
     1921 aaaatgttcc aatatttcca ttggagatga ggttcaattt gaaattagca taacttcaaa
     1981 taagtgtcca aaaaaggatt ctgacagctt taaaattagg cctctgggct ttacggagga
     2041 agtagaggtt attcttcagt acatctgtga atgtgaatgc caaagcgaag gcatccctga
     2101 aagtcccaag tgtcatgaag gaaatgggac atttgagtgt ggcgcgtgca ggtgcaatga
     2161 agggcgtgtt ggtagacatt gtgaatgcag cacagatgaa gttaacagtg aagacatgga
     2221 tgcttactgc aggaaagaaa acagttcaga aatctgcagt aacaatggag agtgcgtctg
     2281 cggacagtgt gtttgtagga agagggataa tacaaatgaa atttattctg gcaaattctg
     2341 cgagtgtgat aatttcaact gtgatagatc caatggctta atttgtggag gaaatggtgt
     2401 ttgcaagtgt cgtgtgtgtg agtgcaaccc caactacact ggcagtgcat gtgactgttc
     2461 tttggatact agtacttgtg aagccagcaa cggacagatc tgcaatggcc ggggcatctg
     2521 cgagtgtggt gtctgtaagt gtacagatcc gaagtttcaa gggcaaacgt gtgagatgtg
     2581 tcagacctgc cttggtgtct gtgctgagca taaagaatgt gttcagtgca gagccttcaa
     2641 taaaggagaa aagaaagaca catgcacaca ggaatgttcc tattttaaca ttaccaaggt
     2701 agaaagtcgg gacaaattac cccagccggt ccaacctgat cctgtgtccc attgtaagga
     2761 gaaggatgtt gacgactgtt ggttctattt tacgtattca gtgaatggga acaacgaggt
     2821 catggttcat gttgtggaga atccagagtg tcccactggt ccagacatca ttccaattgt
     2881 agctggtgtg gttgctggaa ttgttcttat tggccttgca ttactgctga tatggaagct
     2941 tttaatgata attcatgaca gaagggagtt tgctaaattt gaaaaggaga aaatgaatgc
     3001 caaatgggac acgggtgaaa atcctattta taagagtgcc gtaacaactg tggtcaatcc
     3061 gaagtatgag ggaaaactcg aggactacaa agaccatgac ggtgattata aagatcatga
     3121 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     3181 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3241 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3301 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3361 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3421 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3481 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3541 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3601 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3661 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3721 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     3781 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3841 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3901 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3961 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4021 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     4081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4201 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     4261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4441 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4501 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4621 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4681 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4741 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4801 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4861 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4921 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4981 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5041 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5101 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5161 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5221 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5281 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5341 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5401 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5461 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5521 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5581 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5641 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5701 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5761 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5821 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5881 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5941 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6001 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6061 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6121 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6181 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6241 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6301 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6361 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6421 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6481 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6541 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6601 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     6661 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6721 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6781 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6841 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6901 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6961 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7021 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7081 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7141 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7201 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7261 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7321 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7381 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7441 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7501 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7561 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7621 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7681 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7741 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7801 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7861 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7921 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7981 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8041 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8101 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8161 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     8221 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8281 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8341 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
