LOCUS       V045771                 9141 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045771
VERSION     V045771
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..3821
                     /label="Itga3(NM_001306071)"
                     /note="Itga3(NM_001306071)"
                     /gene="Itga3"
     CDS             3828..3893
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3917..4505
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4571..4619
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4821..5249
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5263..5592
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5659..6453
                     /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    6629..6750
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6799..6815)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6823..6839
                     /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(6847..6877)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6892..6913
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7201..7789)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7960..8820)
                     /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(8821..8925)
                     /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 ccaccatgtc ctatctccag acattggtat
      781 ggagcccatg ttcagagagt gtggatttgc aggctgactg gagggaggct tgcaaagcca
      841 ggctgaccca cccgccttcc ttttccagcc ttctggccgg ggctccccgg gacctcgctg
      901 tgggtgatga ctataccaac cggactggtg ctgtgtacct gtgtcccctc acggcccaca
      961 aggacgactg tgaacggatg gacatttcag agaaaagtga ccctgaccat cacattattg
     1021 aggatatgtg gcttggagtg actgtggcca gccagggccc tgcaggtaga gtcctggtct
     1081 gtgcccatcg gtacaccaag gtgctgtggt ctgggctaga agaccagcgg cgcatggtgg
     1141 gcaagtgcta tgtgcgtggc aatgacctac agctcgaccc cggcgatgac tggcagacat
     1201 accacaacga gatgtgtaac agcaacactg actacttgca gaccggcatg tgccagctgg
     1261 gcaccagcgg cggcttcacc cagaacaccg tatacttcgg tgcccctggt gcctacaact
     1321 ggaaaggaaa cagctacatg attcagcgga aggactggga tttatctgaa tatagctaca
     1381 ggggctcaga ggagcaagga aacctttata ttgggtacac ggtgcaggta ggcaacgcca
     1441 tcctacatcc cacggacatc atcactgttg tgacgggtgc cccacggcac caacatatgg
     1501 gcgctgtctt cttgctgaag caagagtcag gtggagacct gcagaggaag caggtgctga
     1561 agggcacgca ggtgggcgct tattttggca gtgccattgc cctggcagac ctgaacaatg
     1621 atgggtggca ggacctcctg gtgggtgctc cctactactt cgaacggaaa gaggaggtag
     1681 ggggtgccgt ctatgtcttc atgaaccagg cgggcgcatc cttccctgat caaccttccc
     1741 tcctgcttca cggccccagc cgctctgcct ttggcatctc tatagccagc atcggtgaca
     1801 tcaaccagga tggattccag gacatcgctg tgggagcccc atttgagggc ttgggcaaag
     1861 tctacatcta ccacagcagc tccggggggc tcctcaggca gccccagcag ataatccatg
     1921 gagagaaact cggactgcca ggcttggcca ccttcggcta ctccctgagc gggaagatgg
     1981 atgtggatga aaacctttac ccagacctgc tagtagggag cctgtcggac cacatcgtgc
     2041 tgctgcgggc ccggcctgtc atcaatatcc tccataggac cttggtggcc aggccagctg
     2101 tgttggaccc tgcgctttgt acagctacct cctgtgttca ggtggagctg tgttttgcct
     2161 acaaccagag cgctgggaac cccaactaca ggcggaacat caccctggct tacacactgg
     2221 aggctgacag ggaccgacgc ccacccaggc tccgatttgc ccgcagccag tcatctgtct
     2281 tccacggctt cttctccatg ccagagacac attgccagac actggagtta ctgctgatgg
     2341 acaatgttcg cgataaactc cgtcctatcg tcattgccat gaactactcc ttacctctgc
     2401 gcatgcctga tcgcctcaag ctcggcttgc ggtctctgga tgcctaccca gtcctcaacc
     2461 aggcacaggc tatggagaat cacactgagg tccacttcca gaaagagtgc gggccagaca
     2521 acaagtgcga cagcaacctg cagatgcgag cagccttcct gtctgagcag ctgcagcctc
     2581 taagcaggct ccagtacagc agagacacta agaaactgtt tttgagcatc aatgtgacca
     2641 attcaccgag cagccaacgg gctggggaag atgcgcatga ggcattgctc accctagagg
     2701 tgccatccgc cctgctactg tcctccgtgc gcccgtctgg gacctgccaa gctaataacg
     2761 agaccatcct gtgtgagcta ggaaacccct tcaaacggaa ccagaggatg gagctgctca
     2821 ttgcctttga ggtcatcggg gtaacgttgc acacgaggga cctcccagta cttcttcagc
     2881 tgtccacgtc gagtcaccag gacaacttgc agcccgtact cctgactttg caggtggact
     2941 acaccctcca ggcctcgctc agcttaatga atcatcggtt gcagagcttc tttggtggca
     3001 cagtgatggg tgaggctgcc atgaaaactg cggaggatgt gggaagtccc ctgaaatatg
     3061 aattccaggt gagcccagtg ggagatgggc tggcagcctt gggcacactg gttctaggtc
     3121 tggagtggcc ctatgaagtt accaatggca agtggctgct gtaccccacg gagatcacca
     3181 tccatagcaa tggttcctgg ccctgccagc cgtctggaaa ccttgtcaac cctctcaacc
     3241 ttactctctc tgaccctgga gtcacaccac tgtcgccaca gcgccgccgg cggcaactgg
     3301 atccaggggg agaccagagt tccccgcctg tcacactagc tgctgccaaa aaagccaagt
     3361 ctgagactgt gttgacctgc tccaatggcc gtgcccgctg tgtgtggctg gagtgccccc
     3421 ttccagacac ctccaacatt accaatgtga ccgtgaaagc acgggtgtgg aacagcacct
     3481 tcattgagga ctacaaagac tttgacagag tcagggtaga tggctgggct accctgttcc
     3541 tgagaaccag catccctacc atcaacatgg agaacaagac cacatggttc tctgtggaca
     3601 ttgactcaga gctggtggag gagctgccgg ctgagattga gctgtggttg gtgcttgtgg
     3661 ccgtgggtgc tgggttgctg ctgctggggc tcatcatcct cctcttgtgg aagtgcggct
     3721 tcttcaagcg agcccgcact cgtgccctgt atgaagctaa gaggcagaag gctgagatga
     3781 agagccagcc gtcagagaca gaaaggctga ccgacgacta cggatccgac tacaaagacc
     3841 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     3901 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3961 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4021 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4081 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4141 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4201 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4261 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     4321 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4381 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4441 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4501 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4561 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4621 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4681 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4741 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     4801 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4861 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4921 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     4981 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5041 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5101 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5161 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     5221 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5521 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5581 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5641 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5701 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5761 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5821 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5881 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5941 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6001 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6061 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6121 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6181 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6241 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6301 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6361 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6421 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6481 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6541 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6601 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6661 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6721 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6781 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7381 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7621 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8941 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     9001 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     9061 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     9121 tgcatgaaga atctgcttag g
//
