LOCUS       sequence-170452-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  AUTHORS   Gonzalez-Agosti C, Wiederhold T, Herndon ME, Gusella J, Ramesh V
  TITLE     Interdomain interaction of merlin isoforms and its influence on 
            intermolecular binding to NHE-RF.
  JOURNAL   J Biol Chem. 1999 Nov 26. 274(48):34438-42.
  PUBMED    10567424
REFERENCE   2  (bases 1 to 6738)
  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"; journalName: "J Biol 
            Chem. 1999 Nov 26. 274(48):34438-42."
COMMENT     Sequence Label: pGEX2T merlin 1-595
FEATURES             Location/Qualifiers
     source          1..6738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2606..2628)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2773..2791)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2859..2963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2964..3032,3033..3824)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3182..3201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3995..4583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4484..4503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4737..4754
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4827..4904
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4905..5987
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4924..4943)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6036..6041,6042..6059,6060..6066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6074..6090
                     /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)."
     primer_bind     6098..6114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6098..6114
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6110..6637
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6130..6147)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6130..6146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6139..6161)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccatggccg gggccatcgc ttcccgcatg
      781 agcttcagct ctctcaagag gaagcaaccc aagacgttca ccgtgaggat cgtcaccatg
      841 gacgccgaga tggagttcaa ttgcgagatg aagtggaaag ggaaggacct ctttgatttg
      901 gtgtgccgga ctctggggct ccgagaaacc tggttctttg gactgcagta cacaatcaag
      961 gacacagtgg cctggctcaa aatggacaag aaggtactgg atcatgatgt ttcaaaggaa
     1021 gaaccagtca cctttcactt cttggccaaa ttttatcctg agaatgctga agaggagctg
     1081 gttcaggaga tcacacaaca tttattcttc ttacaggtaa agaagcagat tttagatgaa
     1141 aagatctact gccctcctga ggcttctgtg ctcctggctt cttacgccgt ccaggccaag
     1201 tatggtgact acgaccccag tgttcacaag cggggatttt tggcccaaga ggaattgctt
     1261 ccaaaaaggg taataaatct gtatcagatg actccggaaa tgtgggagga gagaattact
     1321 gtttggtacg cagagcaccg aggccgagcc agggatgaag ctgaaatgga atatctgaag
     1381 atagctcagg acctggagat gtacggtgtg aactactttg caatccggaa taaaaagggc
     1441 acagagctgc tgcttggagt ggatgccctg gggcttcaca tttatgaccc tgagaacaga
     1501 ctgaccccca agatctcctt cccgtggaat gaaatccgaa acatctcgta cagtgacaag
     1561 gagtttacta ttaaaccact ggataagaaa attgatgtct tcaagtttaa ctcctcaaag
     1621 cttcgtgtta ataagctgat tctccagcta tgtatcgaga accatgatct atttatgagg
     1681 agaaggaaag ccgattcttt ggaagttcag cagatgaaag cccaggccag ggaggagaag
     1741 gctagaaagc agatggagcg gcagcgcctc gctcgagaga agcagatgag ggaggaggct
     1801 gaacgcacga gggatgagtt ggagaggagg ctgctgcaga tgaaagaaga agcaacaatg
     1861 gccaacgaag cactgatgcg gtctgaggag acagctgacc tgttggctga aaaggcccag
     1921 atcaccgagg aggaggcaaa acttctggcc cagaaggccg cagaggctga gcaggaaatg
     1981 cagcgcatca aggccacagc gattcgcacg gaggaggaga agcgcctgat ggagcagaag
     2041 gtgctggaag ccgaggtgct ggcactgaag atggctgagg agtcagagag gagggccaaa
     2101 gaggcagatc agctgaagca ggacctgcag gaagcacgcg aggcggagcg aagagccaag
     2161 cagaagctcc tggagattgc caccaagccc acgtacccgc ccatgaaccc aattccagca
     2221 ccgttgcctc ctgacatacc aagcttcaac ctcattggtg acagcctgtc tttcgacttc
     2281 aaagatactg acatgaagcg gctttccatg gagatagaga aagaaaaagt ggaatacatg
     2341 gaaaagagca agcatctgca ggagcagctc aatgaactca agacagaaat cgaggccttg
     2401 aaactgaaag agagggagac agctctggat attctgcaca atgagaactc cgacagaggt
     2461 ggcagcagca agcacaatac cattaaaaag ctcaccttgc agagcgccaa gtcccgagtg
     2521 gccttctttg aagagctcta gcaggtggaa ttcatcgtga ctgactgacg atctgcctcg
     2581 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2641 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2701 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     2761 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2821 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2941 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3001 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3061 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3181 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     3241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3481 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     3541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3661 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4081 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4141 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4201 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4261 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4321 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4381 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4441 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4501 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4561 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4621 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4681 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4741 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4801 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
     4861 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     4921 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     4981 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5041 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5101 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5161 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5221 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5281 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5341 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5401 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5461 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     5521 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     5581 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     5641 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     5701 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     5761 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     5821 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     5881 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5941 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6001 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6061 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6121 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     6181 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     6241 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     6301 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     6361 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     6421 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     6481 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     6541 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     6601 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     6661 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     6721 atattctgaa atgagctg
//
