LOCUS       sequence-177606-        7610 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 7610)
  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 7610)
  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-hSyntenin2a
FEATURES             Location/Qualifiers
     source          1..7610
                     /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             1632..1652
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1668..2009
                     /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      3368..3555
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3423..3447)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3676..4770)
                     /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(4771..5176)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5218..5234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5218..5234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5231..5253)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5242..5258
                     /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(5266..5272,5273..5290,5291..5296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5311..5332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5449..5466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5620..6208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5700..5719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6379..7170,7171..7239))
                     /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     7002..7021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7240..7344)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7412..7430
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7468..7490)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7590..7609
                     /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 gctttgcaaa gatggataaa gcggaattaa
     1621 ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg taccgccgcc aattttaatc
     1681 aaagtgggaa tattgctgat agctcattgt ccttcacttt cactaacagt agcaacggtc
     1741 cgaacctcat aacaactcaa acaaattctc aagcgctttc acaaccaatt gcctcctcta
     1801 acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat gatggtaata
     1861 attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg tttggaatca
     1921 ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta ttcgatgatg
     1981 aagatacccc accaaaccca aaaaaagaga tctctcgagg atccgaattc atgtcatccc
     2041 tgtacccatc tctagaggac ctaaaagtgg accaagccat tcaggcccag gtcagagcct
     2101 cacccaagat gccagccctg ccagtccagg caacagccat ttccccacca ccagttttgt
     2161 acccaaactt ggcagaactg gaaaattata tgggtctttc cctctccagc caagaagtcc
     2221 aggagagcct gcttcatatt ccagagggtg acagtacagc ggtctcgggc cccgggcccg
     2281 gccagatggt ggcaccggta accgggtaca gcctgggcgt gcggcgagct gagatcaagc
     2341 ccggggtgcg cgagatccac ctgtgcaagg acgagcgcgg caagaccggg ctgaggctgc
     2401 ggaaggtcga ccaggggctc tttgtgcagt tggtccaggc caacacccct gcatcccttg
     2461 tggggctgcg ctttggggac cagctcctgc agattgacgg gcgtgactgt gctgggtgga
     2521 gctcgcacaa agcccatcag gtggtgaaga aggcatcagg cgataagatt gtcatggtgg
     2581 ttcgggacag gccgttccag cggactgtca ccatgcacaa ggacagcatg ggccatgtcg
     2641 gcttcgtgat caagaagggg aagattgtct ctctggtcaa agggagttct gcggcccgca
     2701 acgggctcct caccaaccac tacgtgtgtg aggtggacgg gcagaatgtt atcgggctga
     2761 aggacaaaaa gatcatggag attctggcca cggctgggaa cgttgtcacc ctgaccatca
     2821 tccccagtgt gatctacgag cacatggtca aaaagttgcc tccagtcctg ctccaccaca
     2881 ccatggacca ctccatccca gatgcctgaa gccactgcag ggcagggcag gcaggggggg
     2941 cttcccgccc tcctgcagca aagggcaacc accctcggat gaattccaga tctatgaatc
     3001 gtagatactg aaaaaccccg caagttcact tcaactgtgc atcgtgcacc atctcaattt
     3061 ctttcattta tacatcgttt tgccttcttt tatgtaacta tactcctcta agtttcaatc
     3121 ttggccatgt aacctctgat ctatagaatt ttttaaatga ctagaattaa tgcccatctt
     3181 ttttttggac ctaaattctt catgaaaata tattacgagg gcttattcag aagctttgga
     3241 cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt ctaccttgcc
     3301 agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg ttgacacttc
     3361 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     3421 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     3481 taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga
     3541 ccacacctct accggcatgc cggtcgaaat tcccctaccc tatgaacata ttccattttg
     3601 taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca aatatcataa
     3661 aaaaagagaa tctttttaag caaggatttt cttaacttct tcggcgacag catcaccgac
     3721 ttcggtggta ctgttggaac cacctaaatc accagttctg atacctgcat ccaaaacctt
     3781 tttaactgca tcttcaatgg ccttaccttc ttcaggcaag ttcaatgaca atttcaacat
     3841 cattgcagca gacaagatag tggcgatagg gttgacctta ttctttggca aatctggagc
     3901 agaaccgtgg catggttcgt acaaaccaaa tgcggtgttc ttgtctggca aagaggccaa
     3961 ggacgcagat ggcaacaaac ccaaggaacc tgggataacg gaggcttcat cggagatgat
     4021 atcaccaaac atgttgctgg tgattataat accatttagg tgggttgggt tcttaactag
     4081 gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggaa attcgttctt
     4141 gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat tagctttatc
     4201 caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg ccattcttgt
     4261 gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac catcaccatc
     4321 gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa caacgaagtc
     4381 agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt cggatgcaaa
     4441 gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta gtaaaccttg
     4501 ttcaggtcta acactacctg taccccattt aggaccaccc acagcaccta acaaaacggc
     4561 atcagccttc ttggaggctt ccagcgcctc atctggaagt ggaacacctg tagcatcgat
     4621 agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac gaacatcaga
     4681 aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt ggtcacctgg
     4741 caaaacgacg atcttcttag gggcagacat tagaatggta tatccttgaa atatatatat
     4801 atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta
     4861 ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca
     4921 tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc
     4981 ctttttctcc caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa
     5041 caaaaaattt ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt
     5101 atgttgagga aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct
     5161 gagtattccc acagttgggg gatctcgact ctagctagag gatcaattcg taatcatgtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     7381 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     7441 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     7501 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7561 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
//
