LOCUS       V028052                 9612 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028052
VERSION     V028052
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..4292
                     /label="Itga1(NM_001033228)"
                     /note="Itga1(NM_001033228)"
                     /gene="Itga1"
     CDS             4299..4364
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4388..4976
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5042..5090
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5292..5720
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5734..6063
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6130..6924
                     /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    7100..7221
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7270..7286)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7294..7310
                     /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(7318..7348)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7363..7384
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7672..8260)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8431..9291)
                     /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(9292..9396)
                     /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 ccaccatggt ccccaggcgt cctgctagcc
      781 tagaggtcac tgtagcctgc atctggcttc tcaccgttat cctaggcgtc tgcatctcct
      841 tcaacgttga tgtcaaaaac tccatgagct tcagtggtcc agtggaggac atgtttggat
      901 acaccgttca acagtatgaa aacgaagaag gcaaatgggt tcttattggt tctcctttag
      961 ttggccaacc caaagcaaga actggagatg tctataagtg tccagttggg agagagagat
     1021 caatgccttg tgtgaagttg gatttaccag ttaacacatc gattcctaat gtcacagaaa
     1081 taaaggaaaa catgacattt ggatcaactt tagtcaccaa tccgaaggga ggatttctgg
     1141 catgtgggcc cttgtatgcc tatagatgtg gacatttgca ttatacgact ggcatatgtt
     1201 ctgatgtcag ccctacattt caagttgtga actcgttcgc ccctgtacaa gaatgtagca
     1261 cacagcttga catagtcatc gtcctagatg gctccaacag catctacccc tgggaaagtg
     1321 tcactgcctt tttaaacgac cttcttaaga ggatggacat tggccccaaa cagacacagg
     1381 ttggaattgt acaatatgga gcaaatgtaa cacatgagtt caacctcaat aagtattcat
     1441 ccacagaaga ggtcctggtg gcagcaaaca aaataggccg caggggtggt ctccagacga
     1501 tgacagccct tggaatagac acagccagga aagaggcatt cactgaagct cggggggcca
     1561 gaaggggagt gaaaaaagtc atggttattg tgactgatgg agaatcgcat gacaactatc
     1621 gcctgaaaca ggtcatccaa gactgtgagg atgaaaacat tcagcgattt tccatagcta
     1681 tccttggcca ctataacagg gggaacttaa gcaccgaaaa atttgtggag gagataaaat
     1741 caatcgcaag tgagcccacc gaaaagcact tcttcaacgt ctcagatgag ttggccctgg
     1801 tcactattgt taaagctctg ggagaaagga tatttgcttt ggaagctaca gctgaccagt
     1861 cagcagcttc gtttgaaatg gagatgtctc agaccggctt cagtgctcat tattcacagg
     1921 actgggtcat gcttggagct gttggagcct atgactggaa cggaactgtg gtcatgcaga
     1981 aggctaacca gattgtcatc cctcataaca ccacctttca aactgagccc accaagatga
     2041 acgagcctct tgcttcttac ttgggttaca cagtgaactc tgccaccatc ccgggagatg
     2101 tgctctatat cgctgggcag cctcggtaca atcacaccgg ccaggtcgtc atctacaaga
     2161 tggaggatgg ggacgtcaac attctgcaga cactcagtgg agagcagatc ggatcctact
     2221 ttggtagcgt cttaaccaca attgacatcg acaaagattc ttacactgat ctgcttctcg
     2281 tcggggctcc catgtatatg gggacagaga aagaggaaca gggcaaggtg tacgtgtacg
     2341 ctgtgaatca gacgaggttt gaatatcaaa tgagcctgga accaattaag cagacctgct
     2401 gttcatccct gaaggataat tcatgcacaa aagaaaacaa gaatgagccc tgtggggccc
     2461 gttttggaac agcagttgct gctgtaaaag acctcaacgt ggatggcttt aatgacgtcg
     2521 tgattggagc tcccctggaa gacgaccacg caggagctgt gtacatttat catggcagtg
     2581 gcaagaccat aaggaaagag tatgcgcaac gcattccctc aggtggggat gggaagacgc
     2641 tgaaattttt tggccagtct atccacggag agatggattt aaatggcgat ggtctgactg
     2701 acgtgaccat tggaggcctt ggaggagctg ccctcttctg ggccagagat gtggctgtag
     2761 ttaaagtgac catgaatttt gaacccaaca aagtgaatat tcaaaagaaa aactgccgtg
     2821 tggagggcaa agaaacagta tgcataaatg ctacaatgtg ttttcatgtg aaattgaagt
     2881 ctaaagaaga ctcagtttac gaggctgatc tgcagtaccg tgtcaccctt gattcgctga
     2941 ggcagatatc acggagcttt ttttctggaa ctcaggaaag gaggattcaa agaaacctca
     3001 ccgttcgaga atccgaatgc atcaggcact ccttctacat gttggataag cacgactttc
     3061 aggactccgt gagagtgact ttggatttta acctcactga tccagaaaat gggcccgtgc
     3121 tcgatgacgc tctgccaaac tcagtccatg gacatattcc ttttgccaaa gactgtggga
     3181 acaaggaaag atgcgtttca gacctcaccc tggatgtgtc cacaacagaa aagaacctgc
     3241 tgattgtcag atcccagaat gacaagttca atgtcagcct caccgtcaaa aacaagggag
     3301 acagtgcgta caacacccgg acagtggttc agtattctcc aaatctgatt ttttcaggaa
     3361 ttgaggagat ccaaaaagat agctgcgaat ccaatcaaaa tatcacgtgt agagttggat
     3421 atcctttcct gaggacagga gacgtggtta acttcaaaat aatattccag tttaatacat
     3481 cacatctctc agaaaatgca attattcatt taagtgcaac aagtgacagt gaagagcccc
     3541 tggaatctct ttacgataat gaagtaaata tttccatccc cgtaaaatat gaagtcggac
     3601 tgcagtttta cagttctgcg agtgaacacc acatttcagt tgctgccaat gagactgttc
     3661 ctgagctgat taattccacc aaggacattg gggatgaaat taatgtcttc tacacgatta
     3721 gaaaaagagg gcatttccca atgccagaac ttcggctggc aatttcattt cccaatctga
     3781 cgtcagatgg ctatcctgta ctgtacccaa ctggatggtc atcttctgat aatgtaaact
     3841 gcagacctcg aagccttgag gaccctttgg gtatcaactc tgggaagaaa atgacaatat
     3901 caaagtctga ggttctcaaa agaggcacaa tccaggactg cagtacctgc aagattgcta
     3961 ccatcacgtg tcatctcctt ccctcggatg tgagtcaagt gaatgtctca ctcatcttgt
     4021 ggaaaccaac tttcataaaa gcacattttt ccagcttaaa tcttaccata cggggagaac
     4081 ttcagagtga aaactcatcg ctgactttaa gcagcagcaa ccggaaacga gagctggcta
     4141 ttcagatatc caaagacggg ctcccaggcc gagtgccgct gtgggttatc ctcctgagcg
     4201 cctttgcggg actcctgctg ttaatgctgc ttatattagc tctgtggaag attggattct
     4261 tcaaaaggcc actaaagaag aaaatggaga aaggatccga ctacaaagac catgacggtg
     4321 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     4381 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4441 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4501 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4561 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4621 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4681 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4741 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4801 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4861 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4921 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4981 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5041 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     5101 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5161 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5221 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     5281 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5341 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5401 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     5461 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5521 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5581 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5641 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     5701 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5821 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5881 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5941 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6001 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6061 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     6121 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6301 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6901 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6961 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7021 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7081 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     7141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7201 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     7261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7861 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8101 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9421 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     9481 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9541 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     9601 aatctgctta gg
//
