LOCUS       sequence-250008-        6877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BirA construct in a PCI nls  backbone driving mCherry activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6877)
  AUTHORS   Ling ITC, Sauka-Spengler T
  TITLE     Early chromatin shaping predetermines multipotent vagal neural crest
            into neural, neuronal and mesenchymal lineages.
  JOURNAL   Nat Cell Biol. 2019 Dec;21(12):1504-1517. doi: 
            10.1038/s41556-019-0428-9. Epub 2019 Dec 2.
  PUBMED    31792380
REFERENCE   2  (bases 1 to 6877)
  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: 
            "10.1038/s41556-019-0428-9"; journalName: "Nat Cell Biol"; date: 
            "2019-12"; volume: "21"; issue: "12"; pages: "1504-1517"
COMMENT     Sequence Label: pCI-NLS-BirA-2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          258..1274
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1198..1220
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1282..1301
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1455..1464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1470..1496
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1503..1529
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1536..1562
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1578..1598
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1605..2567
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1605..2567
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2583..2636
                     /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             2643..3353
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2788..2806)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3056..3075
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3480..3499)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3545..3600
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3599..3618)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3991..4007)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3991..4007)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4004..4026)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4015..4031
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4039..4045,4046..4063,4064..4069))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4084..4105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4164..4359
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4210..4345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4365..4499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4402..4421)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4456..4475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4567..4584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4738..5326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4818..4837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5497..6288,6289..6357))
                     /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     6120..6139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6358..6462)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6493..6872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
       61 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
      121 cgcgccaggc ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      181 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      241 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      301 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      361 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      421 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      481 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
      541 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
      601 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
      661 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
      721 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
      781 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
      841 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
      901 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
      961 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1021 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1081 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1141 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1201 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1261 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1321 aagaattgct cgagagttcg aagccttaag tacgcgtagg cgcgccttaa ttaacgttta
     1381 aacgctagct catcgattcg aattcaaggt taagcggccg caagctagat tggcattccg
     1441 gtactgttgg taaagccacc atggccacct atgacgtccc agattacgca agtttgcccg
     1501 ggtatgatgt tcctgattat gctagcctcc cgggttacga tgtgcccgac tatgcctcac
     1561 ttccaggcag atctcctcca aaaaagaaga gaaaggtacg atctatgaag gataacaccg
     1621 tgccactgaa attgattgcc ctgttagcga acggtgaatt tcactctggc gagcagttgg
     1681 gtgaaacgct gggaatgagc cgggcggcta ttaataaaca cattcagaca ctgcgtgact
     1741 ggggcgttga tgtctttacc gttccgggta aaggatacag cctgcctgag cctatccagt
     1801 tacttaatgc taaacagata ttgggtcagc tggatggcgg tagtgtagcc gtgctgccag
     1861 tgattgactc cacgaatcag taccttcttg atcgtatcgg agagcttaaa tcgggcgatg
     1921 cttgcattgc agaataccag caggctggcc gtggtcgccg gggtcggaaa tggttttcgc
     1981 cttttggcgc aaacttatat ttgtcgatgt tctggcgtct ggaacaaggc ccggcggcgg
     2041 cgattggttt aagtctggtt atcggtatcg tgatggcgga agtattacgc aagctgggtg
     2101 cagataaagt tcgtgttaaa tggcctaatg acctctatct gcaggatcgc aagctggcag
     2161 gcattctggt ggagctgact ggcaaaactg gcgatgcggc gcaaatagtc attggagccg
     2221 ggatcaacat ggcaatgcgc cgtgttgaag agagtgtcgt taatcagggg tggatcacgc
     2281 tgcaggaagc ggggatcaat ctcgatcgta atacgttggc ggccatgcta atacgtgaat
     2341 tacgtgctgc gttggaactc ttcgaacaag aaggattggc accttatctg tcgcgctggg
     2401 aaaagctgga taattttatt aatcgcccag tgaaacttat cattggtgat aaagaaatat
     2461 ttggcatttc acgcggaata gacaaacagg gggctttatt acttgagcag gatggaataa
     2521 taaaaccctg gatgggcggt gaaatatccc tgcgtagtgc agaaaaagat atcggatcag
     2581 gagagggcag gggaagtctt ctaacatgcg gggacgtgga ggaaaatccc gggcccagat
     2641 ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2701 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2761 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2821 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2881 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2941 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     3001 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     3061 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     3121 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     3181 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3241 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3301 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc
     3361 gcgactctag aactatagtg agtcgtatta cggccgcaat tcactcctca ggtgcaggct
     3421 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     3481 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     3541 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     3601 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     3661 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     3721 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     3781 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     3841 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     3901 gctgtccctc ttctcttatg gagatccctc gacctgcacc gtcgaccagc tggtcgacgg
     3961 tgcaccgtcg accagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4021 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4081 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4141 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     4381 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4441 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     4501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6481 gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     6541 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6601 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6661 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6721 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6781 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6841 acatctacgt attagtcatc gctattacca tggtcga
//
