LOCUS       sequence-467312-        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Skn7t replaces the zipper and is fused to the IFP-F[1] through the 
            linker (GGGGS) X 2. Plasmid confers ampicillin resistance to 
            DH5-alpha..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  AUTHORS   Tchekanda E, Sivanesan D, Michnick SW
  TITLE     An infrared reporter to detect spatiotemporal dynamics of 
            protein-protein interactions.
  JOURNAL   Nat Methods. 2014 Jun;11(6):641-4. doi: 10.1038/nmeth.2934. Epub 
            2014 Apr 20.
  PUBMED    24747815
REFERENCE   2  (bases 1 to 6742)
  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: 
            "10.1038/nmeth.2934"; journalName: "Nat Methods"; date: "2014-06"; 
            volume: "11"; issue: "6"; pages: "641-4"
COMMENT     Sequence Label: p416GAL1-Skn7t-L-IFP-F[1] (Yeast)
FEATURES             Location/Qualifiers
     source          1..6742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1251..1498
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1267..1285)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1516..1535)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1517..1535)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1545..1562)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1545..1561)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1554..1576)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1702..2157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1789..1808)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1999..2020
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2288..3091)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3092..3307)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3331..3350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3450..3472
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3510..3528)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3569..4072
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4109..4213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4214..4282,4283..5074)
                     /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(4432..4451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5245..5833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5734..5753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5987..6004
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6121..6142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6157..6162,6163..6180,6181..6187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6195..6211
                     /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     6200..6222
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6219..6235
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6219..6235
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6254..6274
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6256..6274
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6298..6739
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6298..6415
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6710..6733
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 aaaccccgga ttctagaatg agcctaacac caaatgctca aaataacaca gtcacgctac
       61 gaaaaggttt ccatgtactg ttggtggaag atgacgcagt gtctatacag ttgtgttcaa
      121 aatttttacg gaaatatggc tgtactgtcc aagttgtcag cgacagtctt tcagctatct
      181 caacactaga gaagtatagg tatgatttgg ttttaatgga cattgttatg ccaaacctag
      241 atggtgccac agcgacatcc attgtcagaa gttttgataa tgagactccc atcattgcca
      301 tgacaggtaa cattatgaat caagacttga tcacatactt acaacatgga atgaatgata
      361 tcttggccaa accattcacg agggatgatt tacactcaat tttaatacgt tatctaaagg
      421 accgtattcc tttatgcgaa cagcaattac cacctcgcaa ctcttcgcca caaactcatt
      481 ccaacaccaa tactgctaat tcgaatccta atacgattaa tgaacagtcg ttagccatgt
      541 taccacaaga taatccgtca actactaccc ctgttacccc aggtgcctct atatcttctg
      601 cacagcatgt tcaacaaggt caacaagaac agcagcatca aattttccat gctcagcagc
      661 agcagcagca tcacaacgcc attgctaatg ctaggtcaga cgtagccata ccgaatttgg
      721 aacatgaaat caacactgta ccacattcct caatgggttc cactccgcaa ttaccacaat
      781 ctacacttca agaaaaccag ctatcaatcg atggtggcgg tggctctgga ggtggtgggt
      841 cctccggagc tcgggaccct ctgccattct ttccacctct gtacctgggc ggccctgaga
      901 ttacaaccga gaactgcgag agagagccta tccacattcc tgggtccatc cagccacacg
      961 gggctctgct cacagctgac ggccactccg gagaggtgct ccaagtgtcc ctgaatgccg
     1021 ctaccttcct gggccaggag cctactgtgc tgcgggggca gaccctggct gccctgctcc
     1081 ccgagcagtg gccagccctg caggcagccc tgcccccagg atgtccagat gccctccaat
     1141 acagggccac cctcgactgg ccagctgctg ggcacctcag cctgactgtg catcgggtgg
     1201 ctgaactcct gatcctggag ttcgaaccta ccgaggcctg gtaactcgag tcatgtaatt
     1261 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     1321 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     1381 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     1441 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     1501 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     1561 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1621 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1681 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1741 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1801 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1861 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1921 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1981 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2041 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2101 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     2161 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact
     2221 gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata
     2281 cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt
     2341 aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa
     2401 taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt
     2461 ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc
     2521 ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa
     2581 tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat gacaattctg
     2641 ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc
     2701 taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc
     2761 tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt
     2821 cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct
     2881 ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta
     2941 atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt
     3001 ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag
     3061 cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg
     3121 ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc
     3181 gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta
     3241 ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg
     3301 aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3361 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3421 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3481 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3541 tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt
     3601 atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg
     3661 tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa
     3721 ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac
     3781 aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca
     3841 ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga
     3901 gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca
     3961 tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt
     4021 tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga
     4081 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4141 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4201 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4261 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4321 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4381 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4441 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4501 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4561 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4621 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4681 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4741 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4801 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4861 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4921 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4981 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5041 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5101 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5161 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5221 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5281 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5341 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5401 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5461 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5521 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5581 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5641 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5701 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5761 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5821 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5881 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5941 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6001 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6061 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6121 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6181 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6241 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tctagtacgg
     6301 attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc gtgcgtcctc
     6361 gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa
     6421 taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc agtaacctgg
     6481 ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata atgcgattag
     6541 ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg atctattaac
     6601 agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac attttcggtt
     6661 tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta atatacctct
     6721 atactttaac gtcaaggaga aa
//
