LOCUS       sequence-75570-e        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding for GFP and the biotinylating enzyme BirA
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  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 6380)
  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-BirA-T2A-eGFP
FEATURES             Location/Qualifiers
     source          1..6380
                     /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"
     CDS             1905..2867
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1905..2867
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2868..2921
                     /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             join(2922..2924,2925..2927,2928..3641)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2922..2924,2925..2927,2928..3638)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2967..2988)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3228..3247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3575..3596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3827..4418
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4484..4501)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4655..5243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4735..4754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5407..6198,6199..6267))
                     /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     6030..6049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6268..6372)
                     /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 ccggatccgc caccatgaag gataacaccg
     1921 tgccactgaa attgattgcc ctgttagcga acggtgaatt tcactctggc gagcagttgg
     1981 gtgaaacgct gggaatgagc cgggcggcta ttaataaaca cattcagaca ctgcgtgact
     2041 ggggcgttga tgtctttacc gttccgggta aaggatacag cctgcctgag cctatccagt
     2101 tacttaatgc taaacagata ttgggtcagc tggatggcgg tagtgtagcc gtgctgccag
     2161 tgattgactc cacgaatcag taccttcttg atcgtatcgg agagcttaaa tcgggcgatg
     2221 cttgcattgc agaataccag caggctggcc gtggtcgccg gggtcggaaa tggttttcgc
     2281 cttttggcgc aaacttatat ttgtcgatgt tctggcgtct ggaacaaggc ccggcggcgg
     2341 cgattggttt aagtctggtt atcggtatcg tgatggcgga agtattacgc aagctgggtg
     2401 cagataaagt tcgtgttaaa tggcctaatg acctctatct gcaggatcgc aagctggcag
     2461 gcattctggt ggagctgact ggcaaaactg gcgatgcggc gcaaatagtc attggagccg
     2521 ggatcaacat ggcaatgcgc cgtgttgaag agagtgtcgt taatcagggg tggatcacgc
     2581 tgcaggaagc ggggatcaat ctcgatcgta atacgttggc ggccatgcta atacgtgaat
     2641 tacgtgctgc gttggaactc ttcgaacaag aaggattggc accttatctg tcgcgctggg
     2701 aaaagctgga taattttatt aatcgcccag tgaaacttat cattggtgat aaagaaatat
     2761 ttggcatttc acgcggaata gacaaacagg gggctttatt acttgagcag gatggaataa
     2821 taaaaccctg gatgggcggt gaaatatcct tgcgtagtgc agaaaaagag ggcagaggaa
     2881 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatggtgagc aagggcgagg
     2941 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3001 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3061 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3121 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3181 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3241 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3301 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3361 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3421 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3481 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3541 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3601 ccgccgggat cactctcggc atggacgagc tgtacaagta aactcgacat ttcgaccggg
     3661 ccgccagcac agtggggggg ggcccgcgat tagtccaatt tgttaaagac aggatatcag
     3721 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     3781 catagataga ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3841 cctgtaggtt tggcaagcta gcttaagtaa gccattttgc aaggcatgga aaaatacata
     3901 actgagaata gagaagttca gatcaaggtt aggaacagag agacaggaga atatgggcca
     3961 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agttggaaca
     4021 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4081 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     4141 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     4201 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     4261 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     4321 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4381 ctgagtgatt gactacccgt cagcgggggt ctttcattct gcattaatga atcggccaac
     4441 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4501 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4561 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4981 gacacgactt atagccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5281 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5341 cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
     5401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6001 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     6061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6361 aggggttccg cgcacatttc
//
