LOCUS       Exported                7106 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses 3xFNLDD in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    3xFNLDD/pMXs-I2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  AUTHORS   Fujita T, Kitaura F, Fujii H
  TITLE     A Critical Role of the Thy28-MYH9 Axis in B Cell-Specific Expression
            of the Pax5 Gene in Chicken B Cells.
  JOURNAL   PLoS One. 2015 Jan 21;10(1):e0116579. doi: 
            10.1371/journal.pone.0116579. eCollection 2015.
  PUBMED    25607658
REFERENCE   2  (bases 1 to 7106)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7106
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1972..2037
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2074..2094
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2107..2715
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     2650..2671
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     misc_feature    2739..3312
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2762..3314
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2906..2923)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3133..3152
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4611..5080
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5210..5227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5381..5969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5461..5480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6133..6993)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6756..6775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6994..7098)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 cccagtgtgg tggtacggga atcgtttagt gaaccgtcag aattaaccat ggactacaaa
     1981 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagctt
     2041 gcggccgcga attcatcgat agatcgatcc cctccaaaaa agaagagaaa ggtcgaacct
     2101 gagggaatga aagcgttaac ggccaggcaa caagaggtgt ttgatctcat ccgtgatcac
     2161 atcagccaga caggtatgcc gccgacgcgt gcggaaatcg cgcagcgttt ggggttccgt
     2221 tccccaaacg cggctgaaga acatctgaag gcgctggcac gcaaaggcgt tattgaaatt
     2281 gtttccggcg catcacgcgg gattcgtctg ttgcaggaag aggaagaagg gttgccgctg
     2341 gtaggtcgtg tggctgccgg tgaaccactt ctggcgcaac agcatattga aggtcattat
     2401 caggtcgatc cttccttatt caagccgaat gctgatttcc tgctgcgcgt cagcgggatg
     2461 tcgatgaaag atatcggcat tatggatggt gacttgctgg cagtgcataa aactcaggat
     2521 gtacgtaacg gtcaggtcgt tgtcgcacgt attgatgacg aggttaccgt taagcgcctg
     2581 aaaaaacagg gcaataaagt cgaactgttg ccagaaaata gcgagtttaa accaattgtc
     2641 gtagatcttc gtcagcagag cttcaccatt gaagggctgg cggttggggt tattcgcaac
     2701 ggcgactggc tgtagtcgag gccgctacgt aaattccgcc cctctccctc ccccccccct
     2761 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2821 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2881 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     2941 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3001 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3061 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3121 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3181 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3241 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3301 acacgatgat aatatggcca caattcctaa gatgagcttt ccatgtaaat ttgtagccag
     3361 cttccttctg attttcaatg tttcttccaa aggtgcagtc tccaaagaga ttacgaatgc
     3421 cttggaaacc tggggtgcct tgggtcagga catcaacttg gacattccta gttttcaaat
     3481 gagtgatgat attgacgata taaaatggga aaaaacttca gacaagaaaa agattgcaca
     3541 attcagaaaa gagaaagaga ctttcaagga aaaagataca tataagctat ttaaaaatgg
     3601 aactctgaaa attaagcatc tgaagaccga tgatcaggat atctacaagg tatcaatata
     3661 tgatacaaaa ggaaaaaatg tgttggaaaa aatatttgat ttgaagattc aagagagggt
     3721 ctcaaaacca aagatctcct ggacttgtat caacacaacc ctgacctgtg aggtaatgaa
     3781 tggaactgac cccgaattaa acctgtatca agatgggaaa catctaaaac tttctcagag
     3841 ggtcatcaca cacaagtgga ccaccagcct gagtgcaaaa ttcaagtgca cagcagggaa
     3901 caaagtcagc aaggaatcca gtgtcgagcc tgtcagctgt ccagagaaag gtctggacat
     3961 ctatctcatc attggcatat gtggaggagg cagcctcttg atggtctttg tggcactgct
     4021 cgttttctat atcaccaaaa ggaaaaaaca gaggagtcgg agaaatgatg aggagctgga
     4081 gacaagagcc cacagagtag ctactgaaga aaggggccgg aagccccaac aaattccagc
     4141 ttcaacccct cagaatccag caacttccca acatcctcct ccaccacctg gtcatcgttc
     4201 ccaggcacct agtcatcgtc ccccgcctcc tggacaccgt gttcagcacc agcctcagaa
     4261 gaggcctcct gctccgtcgg gcacacaagt tcaccagcag aaaggcccgc ccctccccag
     4321 acctcgagtt cagccaaaac ctccccatgg ggcagcagaa aactcattgt ccccttcctc
     4381 taattaaaaa agatagaaac tgtctttttc aataaaaagc actgtggatt tctgcctcta
     4441 gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4501 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4561 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     4621 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4681 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4741 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     4801 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     4861 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4921 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     4981 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     5041 tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat
     5101 taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg
     5161 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5221 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5281 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5821 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6001 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6061 tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat atgagtaaac
     6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7081 acaaataggg gttccgcgca catttc
//
