LOCUS       sequence-177535-        5865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Two-hybrid system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  AUTHORS   Koroll M, Rathjen FG, Volkmer H
  TITLE     The neural cell recognition molecule neurofascin interacts with 
            syntenin-1 but not with syntenin-2, both of which reveal 
            self-associating activity.
  JOURNAL   J Biol Chem. 2001 Apr 6;276(14):10646-54. doi: 
            10.1074/jbc.M010647200. Epub 2001 Jan 4.
  PUBMED    11152476
REFERENCE   2  (bases 1 to 5865)
  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: 
            "10.1074/jbc.M010647200"; journalName: "J Biol Chem"; date: 
            "2001-04-6- 6"; volume: "276"; issue: "14"; pages: "10646-54"
COMMENT     Sequence Label: pGBT9-chNfasc (+RSLE)
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..1178
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1183..1584
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1612..2052
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     terminator      2441..2628
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2496..2520)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2681..3355)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3356..3457)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3471..3487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3471..3487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3484..3506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3495..3511
                     /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(3519..3525,3526..3543,3544..3549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3564..3585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3702..3719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3953..3972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4632..5423,5424..5492))
                     /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     5255..5274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5493..5597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5665..5683
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5721..5743)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5843..5862
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agagtgcacc ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat
       61 atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag
      121 ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa
      181 gttcctattc tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg
      241 ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat
      301 attgcgaata ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt
      361 gctatatccc tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc
      421 gacctctaca ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta
      481 agaactattc atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag
      541 tatatagaga caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa
      601 cgctatcact ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg
      661 gatgccttta tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa
      721 gtggagtcag gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc
      781 ttaacggacc tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag
      841 aaaaaaagta atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta
      901 gaacaaaaaa gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg
      961 ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt
     1021 ttgttttaca aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat
     1081 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     1141 catttttgtt ctacaaaatg aagcacagat gcttcgttgc ttgcatgcaa cttcttttct
     1201 ttttttttct tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat
     1261 gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt
     1321 ccagagctga tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca
     1381 cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata
     1441 aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt
     1501 ttcctcgtca ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa
     1561 gctataccaa gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta
     1621 ctgtcttcta tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa
     1681 gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa
     1741 accaaaaggt ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga
     1801 ctggaacagc tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg
     1861 gattctttac aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat
     1921 aaagatgccg tcacagatag attggcttca gtggagactg atatgcctct aacattgaga
     1981 cagcatagaa taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag
     2041 ttgactgtat cgccggaatt caagaggagc agaggaggga aatacccagt gcgtgacaac
     2101 aaagatgagc acctgaatcc tgaagacaag aacgtagaag atggctcatt cgactacagg
     2161 tctcttgaaa gcgatgaaga caacaaacca ctgcccaaca gtcagacctc cctggatggc
     2221 acgataaagc aacaggagag tgacgacagc ttggtggact acggagaggg tggggaaggg
     2281 cagttcaacg aggacggctc cttcattggc cagtacacag tgaaaaagga caaggaggaa
     2341 acggaaggca acgagagctc ggaagccacg tccccagtca atgctatcta ctcattagcg
     2401 tagcgcaatg caatgggacc tgcagccaag ctaattccgg gcgaatttct tatgatttat
     2461 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     2521 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     2581 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccggcaag
     2641 tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta gcatttttga
     2701 cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat
     2761 caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac
     2821 cagctaacat aaaatgtaag ctttcggggc tctcttgcct tccaacccag tcagaaatcg
     2881 agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg
     2941 aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc
     3001 ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca
     3061 gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat
     3121 tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa
     3181 gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata
     3241 taatacccag caagtcagca tcggaatcta gagcacattc tgcggcctct gtgctctgca
     3301 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     3361 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     3421 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcgtaatcat gtcatagctg
     3481 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3541 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3601 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5641 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5701 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5761 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5821 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactg
//
