LOCUS       Exported                7088 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  Expresses C-terminal V5-tagged BirA biotin ligase.
ACCESSION   .
VERSION     .
KEYWORDS    pEF1a-BirA-V5-neo
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 Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7088
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             18..59
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             69..86
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(109..126)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    115..339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      385..813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(472..491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     682..703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(822..842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        827..1156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1007..1142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1069..1088
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1223..2017
                     /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(1277..1296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1887..1906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2193..2314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2230..2249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2284..2303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2363..2379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2376..2398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2387..2403
                     /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(2411..2441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2456..2477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2594..2611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2754..3342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2834..2853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3513..4373)
                     /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     4136..4155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4374..4478)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4553..4572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4860..6038
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5091..6029
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5986..6006
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     6055..6074
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6055..6073
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6125..7087
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
ORIGIN
        1 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
       61 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
      121 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
      181 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
      241 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
      301 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
      361 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
      421 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
      481 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
      541 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
      601 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
      661 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
      721 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
      781 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
      841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      901 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      961 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     1021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     1081 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     1141 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     1201 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     1261 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     1321 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     1381 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     1441 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     1501 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     1561 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1621 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     1681 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     1741 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     1801 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1861 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     1921 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1981 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     2041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     2161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     2221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     2341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2701 agaacatgtg agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4501 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     4561 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     4621 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     4681 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     4741 gttgacattg attattgact aggcttttgc aaaaagcttt gcaaagatgg ataaagtttt
     4801 aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg
     4861 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     4921 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     4981 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     5041 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     5101 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     5161 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     5221 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     5281 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     5341 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     5401 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     5461 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     5521 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     5581 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     5641 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     5701 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     5761 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     5821 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     5881 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     5941 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     6001 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactaatac
     6061 gactcactat agggagaccc aagctggcta ggtaagcttg gtaccgagct cggatccgcc
     6121 agccatgaag gataacaccg tgccactgaa attgattgcc ctgttagcga acggtgaatt
     6181 tcactctggc gagcagttgg gtgaaacgct gggaatgagc cgggcggcta ttaataaaca
     6241 cattcagaca ctgcgtgact ggggcgttga tgtctttacc gttccgggta aaggatacag
     6301 cctgcctgag cctatccagt tacttaatgc taaacagata ttgggtcagc tggatggcgg
     6361 tagtgtagcc gtgctgccag tgattgactc cacgaatcag taccttcttg atcgtatcgg
     6421 agagcttaaa tcgggcgatg cttgcattgc agaataccag caggctggcc gtggtcgccg
     6481 gggtcggaaa tggttttcgc cttttggcgc aaacttatat ttgtcgatgt tctggcgtct
     6541 ggaacaaggc ccggcggcgg cgattggttt aagtctggtt atcggtatcg tgatggcgga
     6601 agtattacgc aagctgggtg cagataaagt tcgtgttaaa tggcctaatg acctctatct
     6661 gcaggatcgc aagctggcag gcattctggt ggagctgact ggcaaaactg gcgatgcggc
     6721 gcaaatagtc attggagccg ggatcaacat ggcaatgcgc cgtgttgaag agagtgtcgt
     6781 taatcagggg tggatcacgc tgcaggaagc ggggatcaat ctcgatcgta atacgttggc
     6841 ggccatgcta atacgtgaat tacgtgctgc gttggaactc ttcgaacaag aaggattggc
     6901 accttatctg tcgcgctggg aaaagctgga taattttatt aatcgcccag tgaaacttat
     6961 cattggtgat aaagaaatat ttggcatttc acgcggaata gacaaacagg gggctttatt
     7021 acttgagcag gatggaataa taaaaccctg gatgggcggt gaaatatccc tgcgtagtgc
     7081 agaaaaat
//
