LOCUS       sequence-374349-        7696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FL D rerio catenin in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7696)
  AUTHORS   Rangarajan ES, Smith EW, Izard T
  TITLE     The nematode alpha-catenin ortholog, HMP1, has an extended 
            alpha-helix when bound to actin filaments.
  JOURNAL   J Biol Chem. 2023 Feb;299(2):102817. doi: 10.1016/j.jbc.2022.102817.
            Epub 2022 Dec 17.
  PUBMED    36539037
REFERENCE   2  (bases 1 to 7696)
  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."; date: "2023-02"; volume: "299(2):102817. doi"; pages: " 
            10.1016/j.jbc.2022.102817. Epub 2022 Dec 17"
COMMENT     Sequence Label: pGEX-6P--Dr-aCateninFL
FEATURES             Location/Qualifiers
     source          1..7696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3746..3768)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3913..3931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3999..4103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4104..4172,4173..4964)
                     /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(4322..4341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5135..5723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5624..5643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5877..5894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5967..6044
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6045..7127
                     /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(6064..6083)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7176..7181,7182..7199,7200..7206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7214..7230
                     /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     7238..7254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7238..7254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7250..7696,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7270..7287)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7270..7286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7279..7301)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgacgagca
      961 ttaacactgc taacatcaac ttcaaatggg atcccaagag tttggagatc cgtacgcttg
     1021 ctgtggaaag actgctggag ccattggtaa cccaggtcac cacacttgta aactccagca
     1081 acaaaggtcc atctaacaag aagaaggggc ggtctaagaa ggctcacgta ttggcagcct
     1141 cagtggatga agccacccag aacttcttgg agaaaggcga gaagattgct aaagagagcc
     1201 agttcttgaa ggaagagctg accgcagcgg tggaggatgt tcgcaagcaa ggagaatcca
     1261 tgaagatggc ctcgggagag tttgcagagg atccttgttc ttcagtgaaa agaggcaaca
     1321 tggtccgtgc tgcacgtgcc cttctttctg cagtcactca tctgcttgtt ctggctgaca
     1381 tggctgatgt ttaccaactt cttcttcaac tcaaactggt ggaggagaac ctgatgaagg
     1441 tgcgtaatgc tggcactgag caagacctgg gcatccaata caaagctctg aagccagagg
     1501 tggacaaact gaacatgatg gcggccaaga ggcagcagga actcaaggac gtccatcaca
     1561 aagaccaaat ggcagcagct cgcggcgtcc ttcagaggaa cgttcccatg ctgtacacgg
     1621 cttcgcgtgc ctgcctacag cacccagacg tggccgcgta caaggccaac cgggatctga
     1681 tctacaagca gctgcagcac gccgtctctg gcatatctaa tgccgctcag gctaccagca
     1741 gcgaggattc cagcttctcc cagactgcag gtggaggaga gctagcctac gccctcaaca
     1801 actttgataa gcagatcatc gtggacccac tgagtttcag cgaggagaga tttcggccct
     1861 ctcttgagga acgtctggaa agcatcatta gtggcgccgc cctcatggcc gattcctcat
     1921 gcacacgcga cgaccggcgc gaacgcatcg tggctgaatg caactctgtg cgccaggcgc
     1981 tgcaggatct gctctctgag tacatgggca atgctggacg taaggagaag agcgatgccc
     2041 tcaacacggc cattgacaga atgactaaga agacccgtga cctccgcaga cagctgcgga
     2101 aggctgtgat ggaccacgtg tcggactctt ttctggagac caatgttcct ctgctggtat
     2161 tgatcgaagc tgccaagaat ggtaatgaaa aggaggtgaa ggaatatgcc caagttttcc
     2221 gtgagcacgc caacaaactg attgaggttg ccaaccttgc ctgctccatc tccaacaacg
     2281 aagagggagt aaagttggtg cgaatggccg cttcccagct ggaaacactg tgccctcagg
     2341 tgattaatgc agctttggcc ctggctgcca aacctaacag caaggtggct caggataaca
     2401 tggatttgtt taaagaccag tgggaaaaac aggtgcgagt gctcactgac gctgtggatg
     2461 acatcacttc cattgacgac ttcctctgcg tgtctgaaaa tcacattctg gaggatgtga
     2521 acaagtgtgt catcgctctg caagaaaagg atgtggatgg gctcgatcgc actgctggag
     2581 ccattcgggg cagagctgct cgtgtggtcc atttggtcac atctgagatg gacaactata
     2641 tgcctggcgt atacacggag aaggttctgg aagctaccaa acttcttact gaaacagtga
     2701 tgccacgttt caccgagcaa gtagaggcag cagtggaggc tctcagtgcc aacaacgggc
     2761 agccagtgga cgagaacgag ttcattgatg cctcgcgtct ggtgtatgat ggtgtgagag
     2821 acatcaggaa ggcggtgctg atgatcagga ccccagagga actggatgac tcggactttg
     2881 agactgaaga ctttgattct cgcagcagga ccagcgttca gacagaagat gatcagctca
     2941 ttgctggaca gagcgctcgg gccatcatgg ctcagctacc ccaggagcag aaggcgaaga
     3001 ttgcggagca ggtggccagt ttccaggagg agaagagcaa gctggatgca gaggtgtcca
     3061 agtgggatga cagtggaaat gacatcatcg tcttggccaa gcagatgtgc atgatcatga
     3121 tggagatgac agacttcacc agggggaaag gaccactcaa aaacacctca gatgtgatta
     3181 gtgctgccaa gaagattgct gaagccggat caagaatgga caagctgggc cgcgccattg
     3241 cagatcaatg tccagattca gcgtgcaagc aggatctgtt ggcgtatctg cagcgcatcg
     3301 ccctgttctg tcaccagctc aacatctgca gtaaagtcaa agcggaggtg cagaatctgg
     3361 gtggagagct cgtggtgtct ggactggaca gtgccatgtc tctgatccag gcggccaaga
     3421 atctgatgaa ttctgtggtg tccacagtaa aggcctccta cgtggcctcc accaaatacc
     3481 agaaatctca agatatgcaa agcctcaaca tgcccgccat ctcctggaag atgaaagctc
     3541 ccgagaagaa gccgctggtc aaacgcgaga agctggacga cggtcagacc aataaagtca
     3601 agcggtcctc tcaaaagaag cacatcaacc ctgtgcaggc cctgagcgag ttcaaagcta
     3661 tggatagtat ttgaggtcga ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg
     3721 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3781 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3841 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     3901 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3961 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4081 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4141 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4201 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4261 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4321 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4381 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4441 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4501 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4561 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4621 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     4681 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4741 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4801 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4861 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5281 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5881 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     5941 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
     6001 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     6061 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     6121 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     6181 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     6241 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     6301 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     6361 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     6421 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     6481 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     6541 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     6601 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     6661 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     6721 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     6781 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     6841 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     6901 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     6961 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     7021 gaaaaaccac gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7081 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7141 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7201 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7261 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     7321 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     7381 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     7441 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     7501 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     7561 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     7621 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     7681 tgatggcgtt ggaatt
//
