LOCUS       V045283                 6604 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045283
VERSION     V045283
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1338..1358
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman
                     et al., 1985;&nbsp;Kaplan et al., 1988)"
                     /transl_table=1
                     /translation="MGSSKSK"
                     /label="myr"
     CDS             1359..2324
                     /label="Ybx1(NM_011732)"
                     /note="Ybx1(NM_011732)"
                     /gene="Ybx1"
     CDS             2325..2351
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     protein_bind    complement(2370..2394)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             2447..2488
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     polyA_signal    2532..2756
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2826..2859
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(3317..3333)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3807..3911
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3912..4772
                     /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"
     rep_origin      4943..5531
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5819..5840
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5855..5885
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5893..5909
                     /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"
     primer_bind     5917..5933
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6431..6464
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gggagcagca agagcaagat gagcagcgag gccgagaccc
     1381 agcagccgcc cgccgccccc gccgccgccc tcagcgccgc cgacaccaag cccggctcca
     1441 cgggcagcgg cgcgggtagt ggcggcccgg gtggcctcac atcggcggcg cccgccggcg
     1501 gggacaagaa ggtcatcgca acgaaggttt tgggaacagt caaatggttc aatgtaagga
     1561 acggatacgg tttcatcaac aggaatgaca ccaaggaaga cgtatttgta caccagactg
     1621 ccataaagaa gaataacccc aggaagtacc ttcgcagtgt aggcgatgga gagactgtgg
     1681 agtttgatgt tgttgaagga gaaaagggtg cggaggcagc aaatgttaca ggccctggtg
     1741 gagttccagt tcaaggcagt aaatacgcag cagaccgtaa ccattataga cgctatccac
     1801 gtcgtagggg tcctccacgc aattaccagc aaaattacca gaatagtgag agtggggaaa
     1861 agaacgaggg atcggaaagc gctcctgaag gccaggccca acaacgccgg ccctatcgca
     1921 ggcgaaggtt cccaccttac tacatgcgga gaccctatgc gcgtcgacca cagtattcca
     1981 acccccctgt gcaaggagaa gtgatggagg gtgctgacaa ccagggtgca ggagagcaag
     2041 gtagaccagt gagacagaat atgtatcggg gttacagacc acgattccga aggggccctc
     2101 ctcgccaaag acagcctaga gaggatggca atgaagagga caaagaaaat caaggagatg
     2161 agacccaagg tcagcagcca cctcaacgtc ggtatcgccg aaacttcaat taccgacgca
     2221 gacgcccaga gaaccctaaa ccacaagatg gcaaagagac aaaagcagcc gatccaccag
     2281 ctgagaattc gtccgctccc gaggctgagc agggcggggc tgagtaccca tacgacgtcc
     2341 cagactacgc ttaaagggtg ggcgcgccga cccagctttc ttgtacaaag tggttgatat
     2401 ccagcacagt ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     2461 taaccctctc ctcggtctcg attctacgcg taccggttag taatgagttt aaacccgctg
     2521 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2581 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2641 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2701 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2761 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2821 attaaataac ttcgtataat gtatgctata cgaagttata tcgtatgcat gctttgcata
     2881 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
     2941 attccttgaa atacatccac aggtacacct ccaattgact caaatgatgt caattagtct
     3001 atcagaagct tctaaagcca tgacatcatt ttctggaatt ttccaagctg tttaaaggca
     3061 cagtcaactt agtgtatgta aacttctgac ccactggaat tgtgatacag tgaattataa
     3121 gtgaaataat ctgtctgtaa acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag
     3181 atgtcctaac tgacttgcca aaactattgt ttgttaacaa gaaatttgtg gagtagttga
     3241 aaaacgagtt ttaatgactc caacttaagt gtatgtaaac ttccgacttc aactgtaggg
     3301 taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3361 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3421 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     3481 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     3541 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3721 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3781 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5701 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5761 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5821 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5881 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5941 acgccaagct tgcatgcctg caggtcgact ctagacctat acagttgaag tcggaagttt
     6001 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt
     6061 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca
     6121 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc
     6181 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt
     6241 ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat catttgagtc
     6301 aattggaggt gtacctgtgg atgtatttca aggggatcca gacatgataa gatacattga
     6361 tgagtttgga caaaccacaa ctagaatgca gcggccgcta gctcgagatc ttaattaaac
     6421 tagtatcgat ataacttcgt ataatgtatg ctatacgaag ttatatgtac gggccagata
     6481 tacgcgttga cattgattat tgactaggct tttgcaaaaa gctttgcaaa gatggataaa
     6541 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     6601 aatt
//
