LOCUS       sequence-177555-        6479 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 6479)
  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 6479)
  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-hSyntenin2a
FEATURES             Location/Qualifiers
     source          1..6479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..1173
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1178..1579
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1607..2047
                     /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      3050..3237
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3105..3129)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3290..3964)
                     /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(3965..4066)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4080..4096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4080..4096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4093..4115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4104..4120
                     /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(4128..4134,4135..4152,4153..4158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4173..4194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4311..4328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4482..5070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4562..4581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5241..6032,6033..6101))
                     /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     5864..5883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6102..6206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6274..6292
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6330..6352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6452..6471
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaccataac gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata
       61 tacaggcaac acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc
      121 gttgcatttt cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc
      181 tattctctag ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg
      241 aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc
      301 gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat
      361 atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct
      421 ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac
      481 tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat
      541 agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta
      601 tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc
      661 ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga
      721 gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac
      781 ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa
      841 aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca
      901 aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg
      961 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     1021 ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg
     1081 ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt
     1141 ttgttctaca aaatgaagca cagatgcttc gttgcttgca tgcaacttct tttctttttt
     1201 tttcttttct ctctcccccg ttgttgtctc accatatccg caatgacaaa aaaatgatgg
     1261 aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg ttgttccaga
     1321 gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc acgcacacta
     1381 ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg aaataaaaaa
     1441 aagtttgctg tcttgctatc aagtataaat agacctgcaa ttattaatct tttgtttcct
     1501 cgtcattgtt ctcgttccct ttcttccttg tttctttttc tgcacaatat ttcaagctat
     1561 accaagcata caatcaactc caagcttgaa gcaagcctcc tgaaagatga agctactgtc
     1621 ttctatcgaa caagcatgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa
     1681 accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa
     1741 aaggtctccg ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga
     1801 acagctattt ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc
     1861 tttacaggat ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga
     1921 tgccgtcaca gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca
     1981 tagaataagt gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac
     2041 tgtatcgccg gaattcatgt catccctgta cccatctcta gaggacctaa aagtggacca
     2101 agccattcag gcccaggtca gagcctcacc caagatgcca gccctgccag tccaggcaac
     2161 agccatttcc ccaccaccag ttttgtaccc aaacttggca gaactggaaa attatatggg
     2221 tctttccctc tccagccaag aagtccagga gagcctgctt catattccag agggtgacag
     2281 tacagcggtc tcgggccccg ggcccggcca gatggtggca ccggtaaccg ggtacagcct
     2341 gggcgtgcgg cgagctgaga tcaagcccgg ggtgcgcgag atccacctgt gcaaggacga
     2401 gcgcggcaag accgggctga ggctgcggaa ggtcgaccag gggctctttg tgcagttggt
     2461 ccaggccaac acccctgcat cccttgtggg gctgcgcttt ggggaccagc tcctgcagat
     2521 tgacgggcgt gactgtgctg ggtggagctc gcacaaagcc catcaggtgg tgaagaaggc
     2581 atcaggcgat aagattgtca tggtggttcg ggacaggccg ttccagcgga ctgtcaccat
     2641 gcacaaggac agcatgggcc atgtcggctt cgtgatcaag aaggggaaga ttgtctctct
     2701 ggtcaaaggg agttctgcgg cccgcaacgg gctcctcacc aaccactacg tgtgtgaggt
     2761 ggacgggcag aatgttatcg ggctgaagga caaaaagatc atggagattc tggccacggc
     2821 tgggaacgtt gtcaccctga ccatcatccc cagtgtgatc tacgagcaca tggtcaaaaa
     2881 gttgcctcca gtcctgctcc accacaccat ggaccactcc atcccagatg cctgaagcca
     2941 ctgcagggca gggcaggcag ggggggcttc ccgccctcct gcagcaaagg gcaaccaccc
     3001 tcggatgaat tcccggggat ccgtcgacct gcagccaagc taattccggg cgaatttctt
     3061 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     3121 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     3181 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     3241 gccggcaagt gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag
     3301 catttttgac gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc
     3361 cgcttacatc aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg
     3421 tcagtccacc agctaacata aaatgtaagc tttcggggct ctcttgcctt ccaacccagt
     3481 cagaaatcga gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg
     3541 gaataaacga atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa
     3601 atacgagtct tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat
     3661 ctccatgcag ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct
     3721 taggttgatt acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg
     3781 cttttacaag acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg
     3841 gcacacatat aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg
     3901 tgctctgcaa gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag
     3961 acatactcca agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc
     4021 aatgccctcc ctcttggccc tctccttttc ttttttcgac cgaattaatt cgtaatcatg
     4081 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4141 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4201 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4261 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4321 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4381 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4921 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5041 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5101 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5161 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5341 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6241 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6301 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6361 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6421 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagt
//
