LOCUS       sequence-100718-        7185 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Inducible expression of proteins fused to an N-terminal monomeric 
            streptavidin tag (mSA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7185)
  AUTHORS   Beck DB, Narendra V, Drury WJ 3rd, Casey R, Jansen PW, Yuan ZF, 
            Garcia BA, Vermeulen M, Bonasio R
  TITLE     In Vivo Proximity Labeling for the Detection of Protein-Protein and 
            Protein-RNA Interactions.
  JOURNAL   J Proteome Res. 2014 Dec 5;13(12):6135-43. doi: 10.1021/pr500196b. 
            Epub 2014 Oct 29.
  PUBMED    25311790
REFERENCE   2  (bases 1 to 7185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/pr500196b";
            journalName: "J Proteome Res"; date: "2014-12-5- 5"; volume: "13"; 
            issue: "12"; pages: "6135-43"
COMMENT     Sequence Label: pINTO-N-SA
FEATURES             Location/Qualifiers
     source          1..7185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1223..1602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1603..1806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1757..1777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1808..1826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1829..1847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1857..1881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1969..1978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2572..2589)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2578..2802
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2848..3276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2935..2954)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3145..3166
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3285..3305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3290..3619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3470..3605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3532..3551
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3667..3714
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3733..4107
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4237..4358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4274..4293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4328..4347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4407..4423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4407..4423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4420..4442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4431..4447
                     /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(4455..4461,4462..4479,4480..4485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4500..4521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5270..5287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5441..6026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5521..5540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6197..6988,6989..7057))
                     /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     6820..6839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7058..7162)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgttaatt
      181 aagctagcag tctgcaatga gttcgagttg gcgcgcctgt cattctaaat ctctctttca
      241 gcctaaagct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag
      301 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg
      361 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct
      421 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggagg gggctgtccc
      481 cgtgagctcc caggcgcgcc tgtcattcta aatctctctt tcagcctaaa gctttttccc
      541 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
      601 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
      661 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
      721 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccccgtgagc tcccaggcgc
      781 gccaactcgg tactcgggga tgctagcgca ccatcaacga ggaggacatc ttggctaacc
      841 aacccctgtt gttacagtcc atccaggaca gacttggctc gctggtgggc caggacagcg
      901 ggtatgtggg gggtcttccc aagaacgtta aggaaaagct gctgagcttg aagactttac
      961 aaagcgagct attcgaagtt gaaaaggaat ttcaggtgga gatgtttgag ctagagaaca
     1021 agtttctgca gaagtacaag cccatttggg agcagcggtc caggatcatt tcaggacaag
     1081 agcaacccaa gccggaacag atcgcaaagg gccaagagat tgtggagtca ctcaatgaga
     1141 cggagttgca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     1201 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1261 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1321 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1381 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1441 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1501 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1561 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1621 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1681 gacgtcaatg ggagtttgtt ttggaaccaa aatcaacggg actttccaaa atgtcgtaac
     1741 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1801 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
     1861 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     1921 agacaccggg accgatccag cctccggact ctagcgttta aacttaaggc caccatgggc
     1981 gaccccagca aagattctaa agcccaggtg tctgccgccg aagccggaat tacaggcaca
     2041 tggtataatc agctgggatc tacatttatt gtgacagccg gcgccgatgg agccctgaca
     2101 ggaacatatg aatctgccgt gggaaatgcc gaatccagat acaccctgac aggaagatat
     2161 gattctgccc ctgccacaga tggatctgga acagccctgg gatggagagt ggcctggaag
     2221 aataattata gaaacgccca tagcgccaca acatggtctg gccagtatgt gggaggcgcc
     2281 gaagccagaa ttaacacaca gtggaccctg acatctggaa caacagaagc caatgcctgg
     2341 aaaagcactc tgagaggaca tgatacattt acaaaagtga aacctagcgc cgcctctatc
     2401 gatgccgcca agaaggccgg agtgaacaat ggcaatcctc tggatgccgt gcagcagacc
     2461 ggtagcggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatccag
     2521 cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     2581 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2641 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2701 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2761 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2821 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2881 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2941 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3001 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3061 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3121 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3181 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3241 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3301 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3481 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3541 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3601 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3661 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3721 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     3781 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     3841 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     3901 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3961 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     4021 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     4081 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     4141 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4201 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4261 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4321 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4381 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4441 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4501 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4561 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4621 gggcgctctt ccgctagtct gcaatgagtt cgagttggcg cgcctgtcat tctaaatctc
     4681 tctttcagcc taaagctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag
     4741 cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg
     4801 cacgctgccg gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct
     4861 cgctgctgcc ccctagcggg ggagggacgt aattacatcc ctgggggctt tgggaggggg
     4921 ctgtccccgt gagctcccag gcgcgcctgt cattctaaat ctctctttca gcctaaagct
     4981 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     5041 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     5101 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     5161 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc cgtgagctcc
     5221 caggcgcgcc aactcggtac tcggggattt aattaagctc ttccgcttcc tcgctcactg
     5281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5881 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5941 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     6001 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6061 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6121 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6181 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6241 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6301 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6361 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6421 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6481 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6541 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6601 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6661 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6721 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6781 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6841 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6901 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6961 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7021 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7081 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7141 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
