LOCUS       sequence-172438-        6803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal part of the split-BioID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6803)
  AUTHORS   Munter S, Gornemann J, Derua R, Lesage B, Qian J, Heroes E, Waelkens
            E, Eynde AV, Beullens M, Bollen M
  TITLE     Split-BioID: a proximity-biotinylation assay for 
            dimerization-dependent protein interactions.
  JOURNAL   FEBS Lett. 2016 Dec 29. doi: 10.1002/1873-3468.12548.
  PUBMED    28032891
REFERENCE   2  (bases 1 to 6803)
  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"; journalName: "FEBS Lett. 
            2016 Dec 29. doi: 10.1002/1873-3468.12548."
COMMENT     Sequence Label: pcDNA3.1 - PP1g - Flag - BirA* (AA2-140)
FEATURES             Location/Qualifiers
     source          1..6803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(86..415)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      100..235
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(154..173)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     400..420
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(429..857)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     751..770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    903..1127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1116..1133
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1615..1638)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2645..2664)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2646..2664)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2709..2912)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2738..2758)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2913..3292
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3464..3483
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3558..3662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3663..3731,3732..4523)
                     /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     complement(3881..3900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4694..5282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5183..5202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5436..5453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5570..5591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5606..5611,5612..5629,5630..5636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5644..5660
                     /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)."
     primer_bind     5649..5671
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5668..5684
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5668..5684
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5733..5854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5744..5763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5798..5817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6028..6803,1..19))
                     /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
                     VLFVKTDLSGALNELRDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6139..6158)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6749..6768
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatccgaaaa
       61 tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
      121 actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa
      181 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
      241 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
      301 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      361 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      421 acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      481 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
      541 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      601 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
      661 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      721 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      781 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      841 gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt ccgcctcaga
      901 agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt
      961 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
     1021 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
     1081 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca
     1141 gcgctagagt ttgtgctcca gtctccagaa catggacagg tacaggttgg ctccgaatgg
     1201 gctgaaccac ttccgtcctc tgcctcctct gccagcctgc tggtactcgg cgatgcaggc
     1261 gtcgccggac ttcagctctc cgattctgtc cagcaggtac tgattggtgg agtcgatcac
     1321 aggcaacacg gccacgcttc cgccatccag ctgtcccagg atctgcttgg cgttcagcag
     1381 ctggataggc tcaggcaggc tgtagccctt gccaggcacg gtgaacacgt ccactcccca
     1441 gtctctcagt gtctggatgt gcttgttgat ggcggctctg ctcattccca gggtctctcc
     1501 cagctgctcg ccagagtgga actcgccgtt ggccagcagg gcgatcagct tcaggggcac
     1561 ggtgttgtcc ttggatccga attcgaatcc ggagacgtac gaccggttaa cactcttgtc
     1621 gtcatcgtcc ttgtagtcca taacaggcct taagcgtagc tttgcttgct ttgtgatcat
     1681 accccgtggc ggtgtgacag gtctcgtggc attgggcttc tttttctctg caggctttaa
     1741 aatctggaag gagcacatga gggtctcatc cacactcatc atggcgcccg cattgtcaaa
     1801 ctcgccacag tagttgggtg cagaaaacag agtgactaac tgcctctttg caaaaaactc
     1861 atatccatct tcaaccacct gatgggctct acatataaga tccaaatcat gcttatggag
     1921 aaattttgca accacttctg caccaaatgt gaaggacact cctctgtcat tttcacccca
     1981 gcctaagaca tctttatcgg ggtcagacca caaaagatca caaagaagac cttgatctgg
     2041 tacatcagtt ggtctcataa ttcgccgaat ctgctccata gattgaagat ctggtgataa
     2101 acctccatga cagcagaata tcttctcgtc cacgatggct gctatcggta agcagttaaa
     2161 acagtctgtg aacgttttcc acagcttaat gttgtatctt ctcttacact catcataaaa
     2221 tccgtagatt ctattgatgc tggcacactc atggttccct ctaagaagaa aaaagttctc
     2281 cggatacttg attttgtagg ccagcaagag gcagatcgtc tctagtgact gtttgcccct
     2341 gtccacatag tccccgagaa acaaatagtt gctttctgga ggaaagccac cgtattcaaa
     2401 cagacggagc aaatcatagt actgcccgtg gatgtcacca catatcttga gtggtgcttc
     2461 aagttctaaa aggataggct gactgaggaa gatctcccga gacttcaagc acagtccccg
     2521 gatttcattc tcctggagct ggacattctt gcctggcttg gaccctctca cttccagcag
     2581 ccgttggatg atactgtcga tgttgagttt atcgatatcc gccgccatta gccagcttgg
     2641 gtctccctat agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt
     2701 agccagagag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2761 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2821 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2881 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     2941 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     3001 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     3061 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     3121 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     3181 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     3241 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta
     3301 tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg
     3361 caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg
     3421 acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt
     3481 gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca
     3541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3661 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3721 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3781 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3841 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3901 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3961 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4021 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4081 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4141 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4201 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4261 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4321 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4381 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4441 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4501 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4561 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4621 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4681 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4741 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4801 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4861 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4921 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4981 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5041 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5101 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5161 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5221 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5281 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5341 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5401 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5461 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5521 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5581 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5641 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     5701 tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac
     5761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5821 atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga
     5881 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     5941 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     6001 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     6061 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     6121 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     6181 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     6241 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt
     6301 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6361 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     6421 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     6481 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     6541 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     6601 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc cgcagttcat tcagggcacc
     6661 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     6721 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     6781 agcggccgga gaacctgcgt gca
//
