LOCUS       sequence-177561-        6154 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 6154)
  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 6154)
  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-hSyntenin2b
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..1180
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1185..1586
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1614..2054
                     /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      2732..2919
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2787..2811)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2972..3646)
                     /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(3647..3748)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3762..3778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3762..3778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3775..3797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3786..3802
                     /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(3810..3816,3817..3834,3835..3840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3855..3876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3993..4010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5956..5974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6012..6034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6134..6153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgagagtgca ccataacgca tttaagcata aacacgcact atgccgttct tctcatgtat
       61 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc
      121 agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg
      181 aagttcctat tctctagcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag
      241 cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa
      301 atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct
      361 gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac
      421 tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag
      481 taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt
      541 agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc
      601 aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg
      661 gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg
      721 aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa
      781 ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg
      841 agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg
      901 tagaacaaaa aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc
      961 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     1021 ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc
     1081 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     1141 tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt gcttgcatgc aacttctttt
     1201 cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa
     1261 atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg
     1321 ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg
     1381 cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa
     1441 taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt
     1501 gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc
     1561 aagctatacc aagcatacaa tcaactccaa gcttgaagca agcctcctga aagatgaagc
     1621 tactgtcttc tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca
     1681 aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca
     1741 aaaccaaaag gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa
     1801 gactggaaca gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa
     1861 tggattcttt acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga
     1921 ataaagatgc cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga
     1981 gacagcatag aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac
     2041 agttgactgt atcgccggaa ttccagatgg tggcaccggt aaccgggtac agcctgggcg
     2101 tgcggcgagc tgagatcaag cccggggtgc gcgagatcca cctgtgcaag gacgagcgcg
     2161 gcaagaccgg gctgaggctg cggaaggtcg accaggggct ctttgtgcag ttggtccagg
     2221 ccaacacccc tgcatccctt gtggggctgc gctttgggga ccagctcctg cagattgacg
     2281 ggcgtgactg tgctgggtgg agctcgcaca aagcccatca ggtggtgaag aaggcatcag
     2341 gcgataagat tgtcgtggtg gttcgggaca ggccgttcca gcggactgtc accatgcaca
     2401 aggacagcat gggccacgtc ggcttcgtga tcaagaaggg gaagattgtc tctctggtca
     2461 aagggagttc tgcggcctgc aacgggctcc tcaccaacca ctacgtgtgt gaggtggacg
     2521 ggcagaatgt tatcgggctg aaggacaaaa agatcatgga gattctggcc acggctggga
     2581 acgttgtcac cctgaccatc atccccagtg tgatctacga gcacatggtc aaaaagttgc
     2641 ctccagtcct gctccaccac accatggacc actccatccc agatgcctga attcccgggg
     2701 atccgtcgac ctgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat
     2761 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     2821 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     2881 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccggcaa gtgcacaaac
     2941 aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg
     3001 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     3061 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     3121 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     3181 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     3241 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     3301 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     3361 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
     3421 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
     3481 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
     3541 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
     3601 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc caagctgcct
     3661 ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct ccctcttggc
     3721 cctctccttt tcttttttcg accgaattaa ttcgtaatca tgtcatagct gtttcctgtg
     3781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5941 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6001 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     6061 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     6121 tggcttaact atgcggcatc agagcagatt gtac
//
