LOCUS       sequence-420886-        6665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Sap30bp+exon(48nt) into N2a FRT rtTA3 
            expression cells (BioID vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6665)
  AUTHORS   Roth JF, Braunschweig U, Wu M, Li JD, Lin ZY, Larsen B, Weatheritt 
            RJ, Gingras AC, Blencowe BJ
  TITLE     Systematic analysis of alternative exon-dependent interactome 
            remodeling reveals multitasking functions of gene regulatory 
            factors.
  JOURNAL   Mol Cell. 2023 Dec 7;83(23):4222-4238.e10. doi: 
            10.1016/j.molcel.2023.10.034.
  PUBMED    38065061
REFERENCE   2  (bases 1 to 6665)
  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.molcel.2023.10"; journalName: "Mol Cell"; date: 
            "2023-12-7- 7"; volume: "83"; issue: "23"; pages: "4222-4238.e10"
COMMENT     Sequence Label: 5'-BirA-FLAG Sap30bp+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(412..459)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     591..610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    743..967
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     956..973
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1015..1039
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1967..1991)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2004..2027)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(2058..2666,2667..2669,2670..3020))
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(3153..3177)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3199..3217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3236..3254
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3273..3291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3310..3328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3527..3546
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3621..3725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3726..3794,3795..4586)
                     /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(3944..3963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4757..5345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5246..5265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5499..5516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5633..5654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5669..5674,5675..5692,5693..5699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5707..5723
                     /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     5712..5734
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5731..5747
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5731..5747
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5796..5917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5807..5826)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5861..5880
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6047..6665,1..404))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(join(6047..6665,1..404))
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc
       61 gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
      121 tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact
      181 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
      241 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
      301 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
      361 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttggccaa ggaagttcct
      421 atactttcta gagaatagga acttcggaat aggaacttct aggtacgtga accatcaccc
      481 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
      541 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
      601 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
      661 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
      721 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
      781 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
      841 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
      901 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
      961 ggcacagtcg aggctgatca gcgggtttaa acgggccctc tagactcgag tcacaccact
     1021 ttgtacaaga aagttgggta tcactgcttg gccttcttca cgatggtgcc cacagcagag
     1081 atgacggtgg tcttggagcc actggcacta gtggtgacag tgaccactgc tggcagggtg
     1141 gctgtggtgg tgaggatggt gggctgtgct attgtcgtca cggggatggc agagtcccac
     1201 ttgctcttcc tcttctgggc atctgcgaca gctgtgctgg cagtactggt gctggtggcc
     1261 gtggcattgg tcgtggtgcc tttcttggtg cccgttacaa actcaatttt ggttcgctcc
     1321 tttttggcct tttctaattt gtccatttca atcttctggg ccttggctaa tgcctcatag
     1381 taggagtctt cagaccagcc gtggggatcg aacatatcct ttgggtagtt ggtgcccagc
     1441 tcatcaatgg cacagaactg gatcagtttc tcgtagatgc tggggttgcg gaactccttc
     1501 ttcctctgga tgatgtagtt catatccatc ccctccttta tcttccgctc gtagagcttc
     1561 tggatcttat cttgtaggtg atttgaacat ctgccaggcg gttctggtgg gatcttgatt
     1621 tcatcgggtg acatgttgcg aaccctttca gaaaaggagg ccactaattc ctggggatcc
     1681 cgcttctctg cttctgtgtt atcctttggg tcatcagcct caggtttttc agtctctgaa
     1741 tcgtcatctt ccgactgctt actgttctcg tcctcctctt cttcataacc atcctcatca
     1801 ccccctgggc gagagaagtc atcttctcca taggcgtcag ataccaaacc acctttctcc
     1861 tcagtcgctc cgcctacagc gtcgactcca gtttcgccat cagattcagg gtctgaatat
     1921 tccgcataaa ttgcaagaga cgacaaaaca tttttcttcc ccgcggtgcc aacttttttg
     1981 tacaaacttg tgggcgcgcc tagtttatcg tcatcgtctt tgtagtctgg cgcgccgccg
     2041 ccggggccag atcccccctt ctcggcgctc cgcaggctga tctcgccgcc catccagggc
     2101 ttgatgatgc cgtcctgctc cagcagcagg gcgccctgct tgtcgatgcc ccggctgatg
     2161 ccgaagatct ccttgtcgcc gatgatcagc ttcacgggcc ggttgatgaa gttgtccagc
     2221 ttctcccacc ggctcaggta gggggccagg ccctcctgct cgaacagctc cagggcggcc
     2281 cgcagctccc ggatcagcat ggcggccagg gtgttccggt ccaggttgat gccggcctcc
     2341 tgcagggtga tccagccctg gttcaccacg ctctcctcca cccgccgcat ggccatgttg
     2401 atgccggcgc cgatcacgat ctgggcggcg tcgccggtct tgccggtcag ctccaccagg
     2461 atgccggcca gcttccggtc ctgcaggtac aggtcgttgg gccacttcac ccgcaccttg
     2521 tcggcgccca gcttccgcag cacctcggcc atcacgatgc cgatcaccag gctcaggccg
     2581 atggcggcgg cggggccctg ctccagccgc cagaacatgc tcaggtacag gttggcgccg
     2641 aaggggctga accacttccg gccccggccg ccccggccgg cctgctggta ctcggcgatg
     2701 caggcgtcgc cgctcttcag ctcgccgatc cggtccagca ggtactggtt ggtgctgtcg
     2761 atcacgggca gcacggccac gctgccgccg tccagctggc ccaggatctg cttggcgttc
     2821 agcagctgga tgggctcggg caggctgtag cccttgccgg gcacggtgaa cacgtccacg
     2881 ccccagtccc gcagggtctg gatgtgcttg ttgatggcgg cccggctcat gcccagggtc
     2941 tcgcccagct gctcgccgct gtggaactcg ccgttggcca gcagggcgat cagcttcagg
     3001 ggcacggtgt tgtccttcat ggtggcgcta gctggatccg agctcggtac caagcttaag
     3061 tttaaacgct agaggatcca gcctccgcgg aggctggatc ggtcccggtg tcttctatgg
     3121 aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg
     3181 cttatatagg gggtaccttc tctatcactg atagggagct cgagacgtgt cgacttctct
     3241 atcactgata gggagacgtt ctagacgtac cttctctatc actgataggg agctcgagac
     3301 gtgtcgactt ctctatcact gatagggaga cgtgcggcca gcttggatcc ctcgagacgc
     3361 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
     3421 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
     3481 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
     3541 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
     3601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3721 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3781 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3841 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3901 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3961 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4021 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4081 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4141 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4201 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4261 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4321 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4381 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4441 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4501 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4561 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4621 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4681 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4741 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4801 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4861 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4921 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4981 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5041 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5101 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5161 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5221 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5281 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5341 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5401 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5461 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5521 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5581 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5641 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5701 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5761 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     5821 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5881 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     5941 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     6001 acctttcata gaaggcggcg gtggaatcga aatctcgtag tacgtgctat tcctttgccc
     6061 tcggacgagt gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg
     6121 tccagacggc cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc
     6181 ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca
     6241 agctctgata gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc
     6301 ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc
     6361 acggcctcca gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc
     6421 tccagtcaat gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat
     6481 ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga
     6541 gagcctgcgc gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat
     6601 ggggatcagc aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg
     6661 gtccg
//
