LOCUS       sequence-420887-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Sap30bp-exon into N2a FRT rtTA3 
            expression cells (BioID vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  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 6617)
  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..6617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    585..706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(622..641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     676..695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(755..771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(755..771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(768..790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    779..795
                     /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(803..809,810..827,828..833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    848..869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(986..1003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1157..1745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1237..1256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1916..2707,2708..2776))
                     /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     2539..2558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2777..2881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2956..2975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3174..3192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3211..3229
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3248..3266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3285..3303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3325..3349
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3482..3832,3833..3835,3836..4444)
                     /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"
     CDS             4475..4498
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4511..4535
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5415..5439)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5481..5498)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5487..5711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5844..5863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5995..6042
                     /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)."
     CDS             join(6050..6617,1..455)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(6050..6617,1..455)
                     /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 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
       61 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
      121 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
      181 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
      241 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
      301 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
      361 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
      421 gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat ttcgattcca
      481 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
      541 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
      601 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      661 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
      721 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      781 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      841 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      901 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      961 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1021 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1081 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1561 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1741 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1861 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1921 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1981 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2041 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2101 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2161 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2221 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2281 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2341 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2401 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2461 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2521 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2581 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2641 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2701 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2761 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2821 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2881 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     2941 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     3001 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     3061 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     3121 cgatgtacgg gccagatata cgcgtctcga gggatccaag ctggccgcac gtctccctat
     3181 cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtacgtcta
     3241 gaacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag
     3301 agaaggtacc ccctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     3361 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggagg
     3421 ctggatcctc tagcgtttaa acttaagctt ggtaccgagc tcggatccag ctagcgccac
     3481 catgaaggac aacaccgtgc ccctgaagct gatcgccctg ctggccaacg gcgagttcca
     3541 cagcggcgag cagctgggcg agaccctggg catgagccgg gccgccatca acaagcacat
     3601 ccagaccctg cgggactggg gcgtggacgt gttcaccgtg cccggcaagg gctacagcct
     3661 gcccgagccc atccagctgc tgaacgccaa gcagatcctg ggccagctgg acggcggcag
     3721 cgtggccgtg ctgcccgtga tcgacagcac caaccagtac ctgctggacc ggatcggcga
     3781 gctgaagagc ggcgacgcct gcatcgccga gtaccagcag gccggccggg gcggccgggg
     3841 ccggaagtgg ttcagcccct tcggcgccaa cctgtacctg agcatgttct ggcggctgga
     3901 gcagggcccc gccgccgcca tcggcctgag cctggtgatc ggcatcgtga tggccgaggt
     3961 gctgcggaag ctgggcgccg acaaggtgcg ggtgaagtgg cccaacgacc tgtacctgca
     4021 ggaccggaag ctggccggca tcctggtgga gctgaccggc aagaccggcg acgccgccca
     4081 gatcgtgatc ggcgccggca tcaacatggc catgcggcgg gtggaggaga gcgtggtgaa
     4141 ccagggctgg atcaccctgc aggaggccgg catcaacctg gaccggaaca ccctggccgc
     4201 catgctgatc cgggagctgc gggccgccct ggagctgttc gagcaggagg gcctggcccc
     4261 ctacctgagc cggtgggaga agctggacaa cttcatcaac cggcccgtga agctgatcat
     4321 cggcgacaag gagatcttcg gcatcagccg gggcatcgac aagcagggcg ccctgctgct
     4381 ggagcaggac ggcatcatca agccctggat gggcggcgag atcagcctgc ggagcgccga
     4441 gaagggggga tctggccccg gcggcggcgc gccagactac aaagacgatg acgataaact
     4501 aggcgcgccc acaagtttgt acaaaaaagt tggcaccgcg gggaagaaaa atgttttgtc
     4561 gtctcttgca atttatgcgg aatattcaga ccctgaatct gatggcgaaa ctggagtcga
     4621 cgctgtaggc ggagcgactg aggagaaagg tggtttggta tctgacgcct atggagaaga
     4681 tgacttctct cgcccagggg gtgatgagga tggttatgaa gaagaggagg acgagaacag
     4741 taagcagtcg gataacacag aagcagagaa gcgggatccc caggaattag tggcctcctt
     4801 ttctgaaagg gttcgcaaca tgtcacccga tgaaatcaag atcccaccag aaccgcctgg
     4861 cagatgttca aatcacctac aagataagat ccagaagctc tacgagcgga agataaagga
     4921 ggggatggat atgaactaca tcatccagag gaagaaggag ttccgcaacc ccagcatcta
     4981 cgagaaactg atccagttct gtgccattga tgagctgggc accaactacc caaaggatat
     5041 gttcgatccc cacggctggt ctgaagactc ctactatgag gcattagcca aggcccagaa
     5101 gattgaaatg gacaaattag aaaaggccaa aaaggagcga accaaaattg agtttgtaac
     5161 gggcaccaag aaaggcacca cgaccaatgc cacggccacc agcaccagta ctgccagcac
     5221 agctgtcgca gatgcccaga agaggaagag caagtgggac tctgccatcc ccgtgacgac
     5281 aatagcacag cccaccatcc tcaccaccac agccaccctg ccagcagtgg tcactgtcac
     5341 cactagtgcc agtggctcca agaccaccgt catctctgct gtgggcacca tcgtgaagaa
     5401 ggccaagcag tgatacccaa ctttcttgta caaagtggtg tgactcgagt ctagagggcc
     5461 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5521 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5581 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5641 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5701 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt
     6001 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga
     6061 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     6121 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     6181 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     6241 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     6301 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     6361 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     6421 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     6481 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     6541 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     6601 cgaggactgc cccgaag
//
