LOCUS       sequence-364401-        7088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminal V5-tagged BirA biotin ligase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7088)
  AUTHORS   Liu X, Zhang Y, Chen Y, Li M, Zhou F, Li K, Cao H, Ni M, Liu Y, Gu 
            Z, Dickerson KE, Xie S, Hon GC, Xuan Z, Zhang MQ, Shao Z, Xu J
  TITLE     In Situ Capture of Chromatin Interactions by Biotinylated dCas9.
  JOURNAL   Cell. 2017 Aug 24;170(5):1028-1043.e19. doi: 
            10.1016/j.cell.2017.08.003.
  PUBMED    28841410
REFERENCE   2  (bases 1 to 7088)
  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.cell.2017.08"; journalName: "Cell"; date: "2017-08-24- 
            24"; volume: "170"; issue: "5"; pages: "1028-1043.e19"
COMMENT     Sequence Label: pEF1a-BirA-V5-neo
FEATURES             Location/Qualifiers
     source          1..7088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..33)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        18..347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     260..279
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             414..1208
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(468..487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1078..1097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1384..1505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1421..1440)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1475..1494
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1554..1570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1554..1570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1567..1589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1578..1594
                     /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(1602..1608,1609..1626,1627..1632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1647..1668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1785..1802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1945..2533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2025..2044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2704..3495,3496..3564))
                     /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     3327..3346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3565..3669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3744..3763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4051..5229
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4282..5220
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5177..5197
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     5246..5265
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5246..5264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5316..6278
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             5316..6278
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             6297..6338
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6348..6365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6388..6405)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6394..6618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6664..7088,1..4)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6751..6770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6961..6982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
       61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
      301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
      361 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
      421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
      481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
      541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
      601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
      661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
      721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
      781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
      841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
      901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
      961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     1021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     1081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     1141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     1201 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     1261 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1321 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1381 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1441 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1501 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     1561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccaggaaccg
     1921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2461 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2521 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2581 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2641 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2701 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2761 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2821 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2881 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2941 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3001 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3061 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3121 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3181 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3241 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3301 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3361 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3421 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3481 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3541 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3601 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3661 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     3721 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3781 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     3841 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     3901 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac taggcttttg
     3961 caaaaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
     4021 accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc
     4081 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
     4141 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4201 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4261 gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt
     4321 acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt
     4381 gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc
     4441 ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc
     4501 tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt
     4561 tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat
     4621 ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag
     4681 cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag
     4741 tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc
     4801 tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc
     4861 ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag
     4921 tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg
     4981 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
     5041 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
     5101 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
     5161 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
     5221 gtgtcgtgag gaattagctt ggtactaata cgactcacta tagggagacc caagctggct
     5281 aggtaagctt ggtaccgagc tcggatccgc cagccatgaa ggataacacc gtgccactga
     5341 aattgattgc cctgttagcg aacggtgaat ttcactctgg cgagcagttg ggtgaaacgc
     5401 tgggaatgag ccgggcggct attaataaac acattcagac actgcgtgac tggggcgttg
     5461 atgtctttac cgttccgggt aaaggataca gcctgcctga gcctatccag ttacttaatg
     5521 ctaaacagat attgggtcag ctggatggcg gtagtgtagc cgtgctgcca gtgattgact
     5581 ccacgaatca gtaccttctt gatcgtatcg gagagcttaa atcgggcgat gcttgcattg
     5641 cagaatacca gcaggctggc cgtggtcgcc ggggtcggaa atggttttcg ccttttggcg
     5701 caaacttata tttgtcgatg ttctggcgtc tggaacaagg cccggcggcg gcgattggtt
     5761 taagtctggt tatcggtatc gtgatggcgg aagtattacg caagctgggt gcagataaag
     5821 ttcgtgttaa atggcctaat gacctctatc tgcaggatcg caagctggca ggcattctgg
     5881 tggagctgac tggcaaaact ggcgatgcgg cgcaaatagt cattggagcc gggatcaaca
     5941 tggcaatgcg ccgtgttgaa gagagtgtcg ttaatcaggg gtggatcacg ctgcaggaag
     6001 cggggatcaa tctcgatcgt aatacgttgg cggccatgct aatacgtgaa ttacgtgctg
     6061 cgttggaact cttcgaacaa gaaggattgg caccttatct gtcgcgctgg gaaaagctgg
     6121 ataattttat taatcgccca gtgaaactta tcattggtga taaagaaata tttggcattt
     6181 cacgcggaat agacaaacag ggggctttat tacttgagca ggatggaata ataaaaccct
     6241 ggatgggcgg tgaaatatcc ctgcgtagtg cagaaaaatc tagagggccc ttcgaaggta
     6301 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc
     6361 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6421 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6481 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6541 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6601 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     6661 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6841 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     7021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7081 ttaacgcg
//
