LOCUS       V022964                 8594 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022964
VERSION     V022964
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"
     CDS             941..4084
                     /label="ITGAV(NM_002210)"
                     /note="ITGAV(NM_002210)"
                     /gene="ITGAV"
     CDS             4091..4156
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4194..4418
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4464..4892
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4906..5235
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5302..6096
                     /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    6270..6391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6440..6456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6464..6480
                     /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(6488..6518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6533..6554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6842..7427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7598..8458)
                     /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(8459..8563)
                     /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 aagcttggta ccgagctcgg atccgccacc atggcttttc cgccgcggcg
      961 acggctgcgc ctcggtcccc gcggcctccc gcttcttctc tcgggactcc tgctacctct
     1021 gtgccgcgcc ttcaacctag acgtggacag tcctgccgag tactctggcc ccgagggaag
     1081 ttacttcggc ttcgccgtgg atttcttcgt gcccagcgcg tcttcccgga tgtttcttct
     1141 cgtgggagct cccaaagcaa acaccaccca gcctgggatt gtggaaggag ggcaggtcct
     1201 caaatgtgac tggtcttcta cccgccggtg ccagccaatt gaatttgatg caacaggcaa
     1261 tagagattat gccaaggatg atccattgga atttaagtcc catcagtggt ttggagcatc
     1321 tgtgaggtcg aaacaggata aaattttggc ctgtgcccca ttgtaccatt ggagaactga
     1381 gatgaaacag gagcgagagc ctgttggaac atgctttctt caagatggaa caaagactgt
     1441 tgagtatgct ccatgtagat cacaagatat tgatgctgat ggacagggat tttgtcaagg
     1501 aggattcagc attgatttta ctaaagctga cagagtactt cttggtggtc ctggtagctt
     1561 ttattggcaa ggtcagctta tttcggatca agtggcagaa atcgtatcta aatacgaccc
     1621 caatgtttac agcatcaagt ataataacca attagcaact cggactgcac aagctatttt
     1681 tgatgacagc tatttgggtt attctgtggc tgtcggagat ttcaatggtg atggcataga
     1741 tgactttgtt tcaggagttc caagagcagc aaggactttg ggaatggttt atatttatga
     1801 tgggaagaac atgtcctcct tatacaattt tactggcgag cagatggctg catatttcgg
     1861 attttctgta gctgccactg acattaatgg agatgattat gcagatgtgt ttattggagc
     1921 acctctcttc atggatcgtg gctctgatgg caaactccaa gaggtggggc aggtctcagt
     1981 gtctctacag agagcttcag gagacttcca gacgacaaag ctgaatggat ttgaggtctt
     2041 tgcacggttt ggcagtgcca tagctccttt gggagatctg gaccaggatg gtttcaatga
     2101 tattgcaatt gctgctccat atgggggtga agataaaaaa ggaattgttt atatcttcaa
     2161 tggaagatca acaggcttga acgcagtccc atctcaaatc cttgaagggc agtgggctgc
     2221 tcgaagcatg ccaccaagct ttggctattc aatgaaagga gccacagata tagacaaaaa
     2281 tggatatcca gacttaattg taggagcttt tggtgtagat cgagctatct tatacagggc
     2341 cagaccagtt atcactgtaa atgctggtct tgaagtgtac cctagcattt taaatcaaga
     2401 caataaaacc tgctcactgc ctggaacagc tctcaaagtt tcctgtttta atgttaggtt
     2461 ctgcttaaag gcagatggca aaggagtact tcccaggaaa cttaatttcc aggtggaact
     2521 tcttttggat aaactcaagc aaaagggagc aattcgacga gcactgtttc tctacagcag
     2581 gtccccaagt cactccaaga acatgactat ttcaaggggg ggactgatgc agtgtgagga
     2641 attgatagcg tatctgcggg atgaatctga atttagagac aaactcactc caattactat
     2701 ttttatggaa tatcggttgg attatagaac agctgctgat acaacaggct tgcaacccat
     2761 tcttaaccag ttcacgcctg ctaacattag tcgacaggct cacattctac ttgactgtgg
     2821 tgaagacaat gtctgtaaac ccaagctgga agtttctgta gatagtgatc aaaagaagat
     2881 ctatattggg gatgacaacc ctctgacatt gattgttaag gctcagaatc aaggagaagg
     2941 tgcctacgaa gctgagctca tcgtttccat tccactgcag gctgatttca tcggggttgt
     3001 ccgaaacaat gaagccttag caagactttc ctgtgcattt aagacagaaa accaaactcg
     3061 ccaggtggta tgtgaccttg gaaacccaat gaaggctgga actcaactct tagctggtct
     3121 tcgtttcagt gtgcaccagc agtcagagat ggatacttct gtgaaatttg acttacaaat
     3181 ccaaagctca aatctatttg acaaagtaag cccagttgta tctcacaaag ttgatcttgc
     3241 tgttttagct gcagttgaga taagaggagt ctcgagtcct gatcatgtct ttcttccgat
     3301 tccaaactgg gagcacaagg agaaccctga gactgaagaa gatgttgggc cagttgttca
     3361 gcacatctat gagctgagaa acaatggtcc aagttcattc agcaaggcaa tgctccatct
     3421 tcagtggcct tacaaatata ataataacac tctgttgtat atccttcatt atgatattga
     3481 tggaccaatg aactgcactt cagatatgga gatcaaccct ttgagaatta agatctcatc
     3541 tttgcaaaca actgaaaaga atgacacggt tgccgggcaa ggtgagcggg accatctcat
     3601 cactaagcgg gatcttgccc tcagtgaagg agatattcac actttgggtt gtggagttgc
     3661 tcagtgcttg aagattgtct gccaagttgg gagattagac agaggaaaga gtgcaatctt
     3721 gtacgtaaag tcattactgt ggactgagac ttttatgaat aaagaaaatc agaatcattc
     3781 ctattctctg aagtcgtctg cttcatttaa tgtcatagag tttccttata agaatcttcc
     3841 aattgaggat atcaccaact ccacattggt taccactaat gtcacctggg gcattcagcc
     3901 agcgcccatg cctgtgcctg tgtgggtgat cattttagca gttctagcag gattgttgct
     3961 actggctgtt ttggtatttg taatgtacag gatgggcttt tttaaacggg tccggccacc
     4021 tcaagaagaa caagaaaggg agcagcttca acctcatgaa aatggtgaag gaaactcaga
     4081 aactctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4141 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     4201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4381 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4441 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4501 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4561 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4621 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4681 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4741 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4801 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4861 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4921 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4981 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5041 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5101 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5161 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5221 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5281 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5341 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5401 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5461 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5521 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5581 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5641 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5701 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5761 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5821 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5881 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5941 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6001 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6061 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6121 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6181 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6241 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6301 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6361 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6421 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     7381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8581 tgccacctga cgtc
//
