LOCUS       sequence-177605-        8765 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 8765)
  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 8765)
  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: pGAD424-rSyntenin1
FEATURES             Location/Qualifiers
     source          1..8765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..1177
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1182..1583
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1629..1649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1665..2006
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     terminator      4520..4707
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4575..4599)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4828..5922)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5923..6328)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(6370..6386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6370..6386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6383..6405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6394..6410
                     /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(6418..6424,6425..6442,6443..6448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6463..6484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6601..6618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6772..7360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6852..6871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7531..8322,8323..8391))
                     /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     8154..8173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8392..8496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8564..8582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8620..8642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8742..8761
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gagtgcacca taacgcattt aagcataaac acgcactatg ccgttcttct catgtatata
       61 tatatacagg caacacgcag atataggtgc gacgtgaaca gtgagctgta tgtgcgcagc
      121 tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc tattccgaag
      181 ttcctattct ctagctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc
      241 tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata
      301 ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg
      361 ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg
      421 acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa
      481 gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt
      541 atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac
      601 gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg
      661 atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag
      721 tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct
      781 taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga
      841 aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag
      901 aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt
      961 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt
     1021 tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt
     1081 tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc
     1141 atttttgttc tacaaaatga agcacagatg cttcgttgct tgcatgcaac ttcttttctt
     1201 tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga caaaaaaatg
     1261 atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat gtcgttgttc
     1321 cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc attcacgcac
     1381 actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa tttgaaataa
     1441 aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta atcttttgtt
     1501 tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca atatttcaag
     1561 ctataccaag catacaatca actccaagct ttgcaaagat ggataaagcg gaattaattc
     1621 ccgagcctcc aaaaaagaag agaaaggtcg aattgggtac cgccgccaat tttaatcaaa
     1681 gtgggaatat tgctgatagc tcattgtcct tcactttcac taacagtagc aacggtccga
     1741 acctcataac aactcaaaca aattctcaag cgctttcaca accaattgcc tcctctaacg
     1801 ttcatgataa cttcatgaat aatgaaatca cggctagtaa aattgatgat ggtaataatt
     1861 caaaaccact gtcacctggt tggacggacc aaactgcgta taacgcgttt ggaatcacta
     1921 cagggatgtt taataccact acaatggatg atgtatataa ctatctattc gatgatgaag
     1981 ataccccacc aaacccaaaa aaagagatct ctcgaggatc cgaattcgcg gccgcgtcga
     2041 cagcggcagg aacatcgcgg tgttcggggc agaaaacacc tgcaaaaatg tctctctatc
     2101 catctcttga agatttgaag gtagacaaag taattcaggc tcaaactgcc tcttctgcta
     2161 atcctgccag ccaggccttt tttttaaatg tggtcgattc tgctgccttt cctccaggtg
     2221 gaaatctcta tcctaaactg tatccagaac tctctcaata catgggactg tctttaagtg
     2281 aagctgaaat atgtgaaaac atgcctgtgg tctctggagc gccaacccaa gggcagttgg
     2341 tagcaaggcc atccagtgtg aactacatgg tagcacctgt gacaggtaac gatgcgggga
     2401 ttaggagagc agagattaag caagggattc gggaagtcat tttgtgtaag gatcaagatg
     2461 gaaaaattgg gctcagactg aagtcagtag ataatggtat atttgttcaa ctagtccagg
     2521 ctaattcccc agcctccttg gttgggctgc gatttgggga ccaggtgctc cagatcaatg
     2581 gtgaaaactg tgccggctgg agctccgata aggcacacaa ggtgctcaag caggcctttg
     2641 gagagaagat taccatgacc attcgtgata gaccctttga acggacagtt accatgcata
     2701 aggacagcag tggacatgtt ggctttatct ttaaaagtgg aaaaatcaca tccatagtga
     2761 aagacagttc tgctgccaga aacggccttc tcactgatca ccacatctgt gagatcaacg
     2821 gacagaacgt cattggcttg aaggatgctc agattgcaga tatactgtca acagctggga
     2881 ctgtggtcac catcacgatc atgcctgcct ttatcttcga acatattatt aaacggatgg
     2941 ctcccagcat tatgaaaagc ctgatggatc acaccattcc tgaagtttaa accttgtaca
     3001 gtgcatcacc gccgaacgct tccagtcttc ctcttcggca acttctctat ttcaccgcac
     3061 atgaagcttt cccggatcca gggagactgt gctgcatgga gacctgtcta tcttccacca
     3121 tggctgggaa cctcactcta tctgagagcg tcttcagatt ttaagatagg tgtagcctta
     3181 tcctggtttt acagatgtga aactctcaaa agattccctg actttctaga cttggttctt
     3241 tactgggaac ctgatgtttt tctataagcc attgtgatta ggatgactgg tgtaggctta
     3301 gctgtgtccg agaactggtc tgctttctcc gagaagtgca cagtgctagg cacgtgcttc
     3361 agttctgtgg catatgtgga ttgtagtaca tcagcatagt acctgtagga tgattgcctt
     3421 tgatcgctac tttttaagtt tgtgtgtgag tgtgtaaaat tccagttaat caacacttcc
     3481 attccacaaa agcattatac agttctgtgt aggtggcaag agactaatga ttgtcactaa
     3541 attctaacta ccttcactta atgtgcttca gttgtcgcct taactacagt aagcatttag
     3601 aataaaagcc aaaatagaat tattgctgcc ttctgaaacc tgagaggtta gctctctatc
     3661 aagctgttct atgctagccc tgagcagctt actggtgtcg cagagctgcc tagctttacc
     3721 aagattttct agtcactgca taaaggaaac gcctaagagt tgccgtattg ctttctgaga
     3781 tacaaattgc tatatatttc atgtaaaagt tgttatacta aataggataa attttggagt
     3841 ctctttttca ctcgtagatt aagtgggtaa tatctaattt ggcacttgag tcctaacatt
     3901 atgtaacttt agctaggttg gggtgggttt agaatgtttc cctagtcatc tcagtttgtt
     3961 ccattttctg actctatcct gcaaacaaaa atggcaatga cactttatcc tgactctttc
     4021 cttcttagtt tgtgcctatc ttattatttt atgtgatatt atcctattaa atgtataaaa
     4081 atattaaatg tcaaaaaaaa aaaaaaaaaa aanaaaaagt cgacgcggcc gcgaattcca
     4141 gatctatgaa tcgtagatac tgaaaaaccc cgcaagttca cttcaactgt gcatcgtgca
     4201 ccatctcaat ttctttcatt tatacatcgt tttgccttct tttatgtaac tatactcctc
     4261 taagtttcaa tcttggccat gtaacctctg atctatagaa ttttttaaat gactagaatt
     4321 aatgcccatc ttttttttgg acctaaattc ttcatgaaaa tatattacga gggcttattc
     4381 agaagctttg gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt
     4441 gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg gtagatacgt
     4501 tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt
     4561 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     4621 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt
     4681 cgctcttatt gaccacacct ctaccggcat gccggtcgaa attcccctac cctatgaaca
     4741 tattccattt tgtaatttcg tgtcgtttct attatgaatt tcatttataa agtttatgta
     4801 caaatatcat aaaaaaagag aatcttttta agcaaggatt ttcttaactt cttcggcgac
     4861 agcatcaccg acttcggtgg tactgttgga accacctaaa tcaccagttc tgatacctgc
     4921 atccaaaacc tttttaactg catcttcaat ggccttacct tcttcaggca agttcaatga
     4981 caatttcaac atcattgcag cagacaagat agtggcgata gggttgacct tattctttgg
     5041 caaatctgga gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt tcttgtctgg
     5101 caaagaggcc aaggacgcag atggcaacaa acccaaggaa cctgggataa cggaggcttc
     5161 atcggagatg atatcaccaa acatgttgct ggtgattata ataccattta ggtgggttgg
     5221 gttcttaact aggatcatgg cggcagaatc aatcaattga tgttgaacct tcaatgtagg
     5281 aaattcgttc ttgatggttt cctccacagt ttttctccat aatcttgaag aggccaaaac
     5341 attagcttta tccaaggacc aaataggcaa tggtggctca tgttgtaggg ccatgaaagc
     5401 ggccattctt gtgattcttt gcacttctgg aacggtgtat tgttcactat cccaagcgac
     5461 accatcacca tcgtcttcct ttctcttacc aaagtaaata cctcccacta attctctgac
     5521 aacaacgaag tcagtacctt tagcaaattg tggcttgatt ggagataagt ctaaaagaga
     5581 gtcggatgca aagttacatg gtcttaagtt ggcgtacaat tgaagttctt tacggatttt
     5641 tagtaaacct tgttcaggtc taacactacc tgtaccccat ttaggaccac ccacagcacc
     5701 taacaaaacg gcatcagcct tcttggaggc ttccagcgcc tcatctggaa gtggaacacc
     5761 tgtagcatcg atagcagcac caccaattaa atgattttcg aaatcgaact tgacattgga
     5821 acgaacatca gaaatagctt taagaacctt aatggcttcg gctgtgattt cttgaccaac
     5881 gtggtcacct ggcaaaacga cgatcttctt aggggcagac attagaatgg tatatccttg
     5941 aaatatatat atatattgct gaaatgtaaa aggtaagaaa agttagaaag taagacgatt
     6001 gctaaccacc tattggaaaa aacaataggt ccttaaataa tattgtcaac ttcaagtatt
     6061 gtgatgcaag catttagtca tgaacgcttc tctattctat atgaaaagcc ggttccggcg
     6121 ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc gtcaggcgac
     6181 ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat agcgcccctg
     6241 tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga actcttgcat
     6301 cttacgatac ctgagtattc ccacagttgg gggatctcga ctctagctag aggatcaatt
     6361 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7201 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7321 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7501 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7561 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7621 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7681 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7741 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7801 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7861 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7921 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7981 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8041 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8101 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8161 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8221 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8281 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8341 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8401 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8461 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8521 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8581 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8641 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8701 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8761 actga
//
