LOCUS       sequence-75568-e        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector endoding for mOrange and mouse CTCF with a short 
            biotinylation substrate (biotag, Kimetal.,2009) fused to its C-term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  AUTHORS   Nakahashi H, Kwon KR, Resch W, Vian L, Dose M, Stavreva D, Hakim O, 
            Pruett N, Nelson S, Yamane A, Qian J, Dubois W, Welsh S, Phair RD, 
            Pugh BF, Lobanenkov V, Hager GL, Casellas R
  TITLE     A genome-wide map of CTCF multivalency redefines the CTCF code.
  JOURNAL   Cell Rep. 2013 May 30;3(5):1678-89. doi: 
            10.1016/j.celrep.2013.04.024. Epub 2013 May 23.
  PUBMED    23707059
REFERENCE   2  (bases 1 to 7703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2013.04.024"; journalName: "Cell Rep"; date: 
            "2013-05-30- 30"; volume: "3"; issue: "5"; pages: "1678-89"
COMMENT     Sequence Label: pMy-MouseCTCFbiotag-T2A-mOrange
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             7..600
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             1090..1506
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1436..1458
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1474..1490
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1516..1890
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1828..1844
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1921..1930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4138..4182
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4201..4254
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4255..4965
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     LTR             5150..5741
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5807..5824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6730..7521,7522..7590))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7353..7372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7591..7695)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcataatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
       61 caaggcatgg aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga
      121 tggaacagct gaatatgggc caaagcggat atctgtggta agcagttcct gccccggctc
      181 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      241 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
      301 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      361 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      421 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
      481 cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc
      541 gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt
      601 tggaggttcc accgagattt ggagacccca gcccagggac caccgacccc cccgccggga
      661 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
      721 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
      781 cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccaggtcg
      841 ggggccgttt ttgtggcccg acctgagtcc aaaaagcccg atcgttttgg actctttggt
      901 gcacccccct tagaggaggg atatgtggtt ctggtaggag acgagaacct aaaacagttc
      961 ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc gcgtcttgtc
     1021 tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
     1081 atgggcccgg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
     1141 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
     1201 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
     1261 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
     1321 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
     1381 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
     1441 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
     1501 ggcgccccca tatggccata tgagatctta tatggggcac ccccgcccct tgtaaacttc
     1561 cctgaccctg acatgacaag agttactaac agcccctctc tccaagctca cttacaggct
     1621 ctctacttag tccagcacga agtctggaga cctctggcgg cagcctacca agaacaactg
     1681 gaccgaccgg tggtacctca cccttaccga gtcggcgaca cagtgtgggt ccgccgacac
     1741 cagactaaga acctagaacc tcgctggaaa ggaccttaca cagtcctgct gaccaccccc
     1801 accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt gaaggctgcc
     1861 gaccccgggg gtggaccatc ctctagactg ccggatccca gtgtggtggt acgggaattc
     1921 gccaccatgg aaggtgaggc ggttgaagcc attgtggagg agtctgaaac tttcattaaa
     1981 ggaaaagaaa gaaagactta ccagagacgc cgggaagggg gccaggaaga agatgcttgc
     2041 cacctgcccc agaaccagac agatgggggt gaggtggtcc aggatgtcaa cagcagtgta
     2101 cagatggtaa tgatggaaca gctggatcct acccttctcc agatgaagac tgaagtcatg
     2161 gagggtacag tggctccgga agcagaggct gcagtggacg atacccagat cataaccttg
     2221 caggttgtaa atatggagga acagcccatt aacataggag agcttcagct tgtccaagta
     2281 cctgttcctg tgacggtacc tgttgctact acttcagtag aagaacttca gggggcttat
     2341 gagaatgaag tgtctaaaga gggccttgca gaaagtgaac cgatgatatg tcacacctta
     2401 cctttgcctg aaggatttca ggtggtgaaa gtgggggcca atggagaagt ggagacacta
     2461 gagcagggcg agcttcctcc tcaggaagac tctagctggc aaaaagaccc agactatcag
     2521 ccaccagcca aaaaaacaaa gaaaaccaaa aagagcaaac ttcgttacac agaagagggc
     2581 aaagacgtgg atgtgtctgt gtatgatttt gaggaagaac agcaggaagg actgctgtct
     2641 gaggttaatg cagagaaagt agttggtaat atgaagcctc cgaagccaac aaaaattaaa
     2701 aaaaaaggtg taaagaaaac attccagtgt gagctttgca gttacacatg tccccggcgt
     2761 tcaaatttgg atcgtcacat gaaaagccac actgatgaga gaccacacaa atgccatctg
     2821 tgtggcagag cattcagaac agtgaccctc ctgaggaatc atctgaacac acacacaggt
     2881 actcgtcctc acaagtgccc agactgcgat atggcctttg tgaccagtgg agaattggtg
     2941 cggcatcgtc gttataaaca cactcatgag aaaccattta agtgttccat gtgtgattat
     3001 gccagtgtag aagtcagcaa attaaaacga cacattcgct ctcatactgg agagcgcccg
     3061 ttccagtgca gtttgtgcag ttatgccagc agggacacat acaagctgaa aaggcatatg
     3121 agaacccatt caggggaaaa accttatgaa tgttatattt gtcacgctcg gtttacccag
     3181 agtggtacca tgaagatgca cattttacag aagcacacag aaaatgtggc caaatttcat
     3241 tgtccccatt gtgacactgt catagcccga aaaagtgatt tgggtgtcca cttgcgaaag
     3301 cagcattcct atattgaaca gggcaaaaaa tgtcgctact gtgatgctgt gtttcatgag
     3361 cgatatgctc tcatccagca tcaaaaatca cacaaaaatg agaagcgctt caagtgtgac
     3421 cagtgtgatt atgcttgtag acaggagcgg cacatgatca tgcacaagcg cactcacacg
     3481 ggggagaagc cttatgcctg cagccactgc gacaagacct tccgccagaa acagctcctc
     3541 gacatgcatt tcaagcgcta tcatgatccc aactttgtcc ctgctgcctt tgtctgttcc
     3601 aagtgtggga aaacattcac ccgccggaac acaatggcaa gacatgcaga taactgtgct
     3661 ggtccagatg gcgtagaggg ggaaaatgga ggggagacaa agaagagcaa acgaggaaga
     3721 aaaagaaaga tgcgatctaa aaaagaagac tcctctgaca gtgaagaaaa tgctgagccg
     3781 gatctcgatg acaatgagga ggaggaggag cctgctgtag aaattgaacc tgagccagag
     3841 cctcagcccc agcctccgcc tccacctcag cctgtggccc cggccccccc acctgccaag
     3901 aagagaaggg ggcgacctcc tggaagaacc aaccagccca aacagaacca gccaacagcc
     3961 atcattcagg tcgaagatca gaatacaggt gcaattgaga acattatagt tgaagtcaaa
     4021 aaggagccag atgccgagcc tgcggagggg gaagaagagg aggctcaggc agccaccaca
     4081 gacgccccca acggagacct cacgcctgag atgatcctca gcatgatgga ccggtccggc
     4141 ctgaacgaca tcttcgaggc tcagaaaatc gaatggcacg aaggcgcgcc gagctcgagg
     4201 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatggtg
     4261 agcaagggcg aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcgc
     4321 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4381 tacgagggct ttcagaccgc taagctgaag gtgaccaagg gtggccccct gcccttcgcc
     4441 tgggacatcc tgtcccctca tttcacctac ggctccaagg cctacgtgaa gcaccccgcc
     4501 gacatccccg actacttcaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     4561 aactacgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     4621 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtgatgcag
     4681 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggtgccctg
     4741 aagggcaaga tcaagatgag gctgaagctg aaggacggcg gccactacac ctccgaggtc
     4801 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacat cgtcgacatc
     4861 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     4921 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaactcga catttcgacc
     4981 gggccgccag cacagtgggg gggggcccgc gattagtcca atttgttaaa gacaggatat
     5041 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     5101 agccatagat agaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     5161 ccacctgtag gtttggcaag ctagcttaag taagccattt tgcaaggcat ggaaaaatac
     5221 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagg agaatatggg
     5281 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagttgga
     5341 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     5401 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     5461 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     5521 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     5581 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     5641 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5701 cctctgagtg attgactacc cgtcagcggg ggtctttcat tctgcattaa tgaatcggcc
     5761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatagcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     6721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6841 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7321 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     7381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7681 aataggggtt ccgcgcacat ttc
//
