LOCUS       sequence-177610-        7285 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 7285)
  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 7285)
  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-hSyntenin2b
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..1184
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1189..1590
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1636..1656
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1672..2013
                     /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      3047..3234
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3102..3126)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3355..4449)
                     /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(4450..4855)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4897..4913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4897..4913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4910..4932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4921..4937
                     /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(4945..4951,4952..4969,4970..4975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4990..5011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5128..5145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5299..5887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5379..5398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6058..6849,6850..6918))
                     /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     6681..6700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6919..7023)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7091..7109
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7147..7169)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(7269..7285,1..3)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtactgagag tgcaccataa cgcatttaag cataaacacg cactatgccg ttcttctcat
       61 gtatatatat atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt
      121 gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat
      181 tccgaagttc ctattctcta gctagaaagt ataggaactt cagagcgctt ttgaaaacca
      241 aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac
      301 taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat
      361 ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga acttgcatct
      421 aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt
      481 gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat
      541 acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat
      601 catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata
      661 atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg
      721 cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt
      781 ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca
      841 aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt
      901 tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca
      961 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     1021 gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg
     1081 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     1141 gcgttgcatt tttgttctac aaaatgaagc acagatgctt cgttgcttgc atgcaacttc
     1201 ttttcttttt ttttcttttc tctctccccc gttgttgtct caccatatcc gcaatgacaa
     1261 aaaaatgatg gaagacacta aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc
     1321 gttgttccag agctgatgag gggtatctcg aagcacacga aactttttcc ttccttcatt
     1381 cacgcacact actctctaat gagcaacggt atacggcctt ccttccagtt acttgaattt
     1441 gaaataaaaa aaagtttgct gtcttgctat caagtataaa tagacctgca attattaatc
     1501 ttttgtttcc tcgtcattgt tctcgttccc tttcttcctt gtttcttttt ctgcacaata
     1561 tttcaagcta taccaagcat acaatcaact ccaagctttg caaagatgga taaagcggaa
     1621 ttaattcccg agcctccaaa aaagaagaga aaggtcgaat tgggtaccgc cgccaatttt
     1681 aatcaaagtg ggaatattgc tgatagctca ttgtccttca ctttcactaa cagtagcaac
     1741 ggtccgaacc tcataacaac tcaaacaaat tctcaagcgc tttcacaacc aattgcctcc
     1801 tctaacgttc atgataactt catgaataat gaaatcacgg ctagtaaaat tgatgatggt
     1861 aataattcaa aaccactgtc acctggttgg acggaccaaa ctgcgtataa cgcgtttgga
     1921 atcactacag ggatgtttaa taccactaca atggatgatg tatataacta tctattcgat
     1981 gatgaagata ccccaccaaa cccaaaaaaa gagatctctc gaggatccga attccagatg
     2041 gtggcaccgg taaccgggta cagcctgggc gtgcggcgag ctgagatcaa gcccggggtg
     2101 cgcgagatcc acctgtgcaa ggacgagcgc ggcaagaccg ggctgaggct gcggaaggtc
     2161 gaccaggggc tctttgtgca gttggtccag gccaacaccc ctgcatccct tgtggggctg
     2221 cgctttgggg accagctcct gcagattgac gggcgtgact gtgctgggtg gagctcgcac
     2281 aaagcccatc aggtggtgaa gaaggcatca ggcgataaga ttgtcgtggt ggttcgggac
     2341 aggccgttcc agcggactgt caccatgcac aaggacagca tgggccacgt cggcttcgtg
     2401 atcaagaagg ggaagattgt ctctctggtc aaagggagtt ctgcggcctg caacgggctc
     2461 ctcaccaacc actacgtgtg tgaggtggac gggcagaatg ttatcgggct gaaggacaaa
     2521 aagatcatgg agattctggc cacggctggg aacgttgtca ccctgaccat catccccagt
     2581 gtgatctacg agcacatggt caaaaagttg cctccagtcc tgctccacca caccatggac
     2641 cactccatcc cagatgcctg aattccagat ctatgaatcg tagatactga aaaaccccgc
     2701 aagttcactt caactgtgca tcgtgcacca tctcaatttc tttcatttat acatcgtttt
     2761 gccttctttt atgtaactat actcctctaa gtttcaatct tggccatgta acctctgatc
     2821 tatagaattt tttaaatgac tagaattaat gcccatcttt tttttggacc taaattcttc
     2881 atgaaaatat attacgaggg cttattcaga agctttggac ttcttcgcca gaggtttggt
     2941 caagtctcca atcaaggttg tcggcttgtc taccttgcca gaaatttacg aaaagatgga
     3001 aaagggtcaa atcgttggta gatacgttgt tgacacttct aaataagcga atttcttatg
     3061 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     3121 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     3181 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc
     3241 ggtcgaaatt cccctaccct atgaacatat tccattttgt aatttcgtgt cgtttctatt
     3301 atgaatttca tttataaagt ttatgtacaa atatcataaa aaaagagaat ctttttaagc
     3361 aaggattttc ttaacttctt cggcgacagc atcaccgact tcggtggtac tgttggaacc
     3421 acctaaatca ccagttctga tacctgcatc caaaaccttt ttaactgcat cttcaatggc
     3481 cttaccttct tcaggcaagt tcaatgacaa tttcaacatc attgcagcag acaagatagt
     3541 ggcgataggg ttgaccttat tctttggcaa atctggagca gaaccgtggc atggttcgta
     3601 caaaccaaat gcggtgttct tgtctggcaa agaggccaag gacgcagatg gcaacaaacc
     3661 caaggaacct gggataacgg aggcttcatc ggagatgata tcaccaaaca tgttgctggt
     3721 gattataata ccatttaggt gggttgggtt cttaactagg atcatggcgg cagaatcaat
     3781 caattgatgt tgaaccttca atgtaggaaa ttcgttcttg atggtttcct ccacagtttt
     3841 tctccataat cttgaagagg ccaaaacatt agctttatcc aaggaccaaa taggcaatgg
     3901 tggctcatgt tgtagggcca tgaaagcggc cattcttgtg attctttgca cttctggaac
     3961 ggtgtattgt tcactatccc aagcgacacc atcaccatcg tcttcctttc tcttaccaaa
     4021 gtaaatacct cccactaatt ctctgacaac aacgaagtca gtacctttag caaattgtgg
     4081 cttgattgga gataagtcta aaagagagtc ggatgcaaag ttacatggtc ttaagttggc
     4141 gtacaattga agttctttac ggatttttag taaaccttgt tcaggtctaa cactacctgt
     4201 accccattta ggaccaccca cagcacctaa caaaacggca tcagccttct tggaggcttc
     4261 cagcgcctca tctggaagtg gaacacctgt agcatcgata gcagcaccac caattaaatg
     4321 attttcgaaa tcgaacttga cattggaacg aacatcagaa atagctttaa gaaccttaat
     4381 ggcttcggct gtgatttctt gaccaacgtg gtcacctggc aaaacgacga tcttcttagg
     4441 ggcagacatt agaatggtat atccttgaaa tatatatata tattgctgaa atgtaaaagg
     4501 taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac aataggtcct
     4561 taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct
     4621 attctatatg aaaagccggt tccggcgctc tcacctttcc tttttctccc aatttttcag
     4681 ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga
     4741 atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg
     4801 gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca cagttggggg
     4861 atctcgactc tagctagagg atcaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     4921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5701 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7021 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7081 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     7141 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7201 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     7261 taactatgcg gcatcagagc agatt
//
