LOCUS       sequence-368775-        7688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human alpha-catenin with an N-terminal GST tag and a 
            PRescission Protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7688)
  AUTHORS   Rangarajan ES, Smith EW, Izard T
  TITLE     Distinct inter-domain interactions of dimeric versus monomeric 
            alpha-catenin link cell junctions to filaments.
  JOURNAL   Commun Biol. 2023 Mar 16;6(1):276. doi: 10.1038/s42003-023-04610-x.
  PUBMED    36928388
REFERENCE   2  (bases 1 to 7688)
  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: "Commun 
            Biol."; date: "2023-03-16"; volume: "6(1):276. doi"; pages: " 
            10.1038/s42003-023-04610-x"
COMMENT     Sequence Label: pGEX-6P-acatenin
FEATURES             Location/Qualifiers
     source          1..7688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(182..187,188..203,204..210)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    218..234
                     /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             257..910
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     868..890
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             917..940
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3737..3759)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3904..3922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3990..4094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4095..4163,4164..4955)
                     /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(4313..4332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5126..5714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5615..5634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5868..5885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5958..6035
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6036..7118
                     /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(6055..6074)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7167..7172,7173..7190,7191..7197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7205..7221
                     /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     7229..7245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7229..7245
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7241..7688,1..80)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7261..7278)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7261..7277)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7270..7292)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
       61 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
      121 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
      181 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
      241 acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca
      301 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
      361 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
      421 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
      481 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
      541 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
      601 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
      661 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
      721 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
      781 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
      841 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
      901 tcctccaaaa tcggatctgg aagttctgtt ccaggggccc ctgggatcca tgactgctgt
      961 ccatgcaggc aacataaact tcaagtggga tcctaaaagt ctagagatca ggactctggc
     1021 agttgagaga ctgttggagc ctcttgttac acaggttaca acccttgtaa acaccaatag
     1081 taaagggccc tctaataaga agagaggtcg ttctaagaag gcccatgttt tggctgcatc
     1141 tgttgaacaa gcaactgaga atttcttgga gaagggggat aaaattgcga aggagagcca
     1201 gtttctcaag gaggagcttg tggctgctgt agaagatgtt cgaaaacaag gtgatttgat
     1261 gaaggctgct gcaggagagt tcgcagatga tccctgctct tctgtgaagc gaggcaacat
     1321 ggttcgggca gctcgagctt tgctctctgc tgttacccgg ttgctgattt tggctgacat
     1381 ggcagatgtc tacaaattac ttgttcagct gaaagttgtg gaagatggta tcttgaagtt
     1441 gaggaatgct ggcaatgaac aagacttagg aatccagtat aaagccctaa aacctgaagt
     1501 ggataagctg aacattatgg cagccaaaag acaacaggaa ttgaaagatg ttggccatcg
     1561 tgatcagatg gctgcagcta gaggaatcct gcagaagaac gttccgatcc tctatactgc
     1621 atcccaggca tgcctacagc accctgatgt cgcagcctat aaggccaaca gggacctgat
     1681 atacaagcag ctgcagcagg cggtcacagg catttccaat gcagcccagg ccactgcctc
     1741 agacgatgcc tcacagcacc agggtggagg aggaggagaa ctggcatatg cactcaataa
     1801 ctttgacaaa caaatcattg tggacccctt gagcttcagc gaggagcgct ttaggccttc
     1861 cctggaggag cgtctggaaa gcatcattag tggggctgcc ttgatggccg actcgtcctg
     1921 cacgcgtgat gaccgtcgtg agcgaattgt ggcagagtgt aatgctgtcc gccaggccct
     1981 gcaggacctg ctttcggagt acatgggcaa tgctggacgt aaagaaagaa gtgatgcact
     2041 caattctgca atagataaaa tgaccaagaa gaccagggac ttgcgtagac agctccgcaa
     2101 agctgtcatg gaccacgttt cagattcttt cctggaaacc aatgttccac ttttggtatt
     2161 gattgaagct gcaaagaatg gaaatgagaa agaagttaag gagtatgccc aagttttccg
     2221 tgaacatgcc aacaaattga ttgaggttgc caacttggcc tgttccatct caaataatga
     2281 agaaggtgta aagcttgttc gaatgtctgc aagccagtta gaagccctct gtcctcaggt
     2341 tattaatgct gcactggctt tagcagcaaa accacagagt aaactggccc aagagaacat
     2401 ggatcttttt aaagaacaat gggaaaaaca agtccgtgtt ctcacagatg ctgtcgatga
     2461 cattacttcc attgatgact tcttggctgt ctcagagaat cacattttgg aagatgtgaa
     2521 caaatgtgtc attgctctcc aagagaagga tgtggatggc ctggaccgca cagctggtgc
     2581 aattcgaggc cgggcagccc gggtcattca cgtagtcacc tcagagatgg acaactatga
     2641 gccaggagtc tacacagaga aggttctgga agccactaag ctgctctcca acacagtcat
     2701 gccacgtttt actgagcaag tagaagcagc cgtggaagcc ctcagctcgg accctgccca
     2761 gcccatggat gagaatgagt ttatcgatgc ttcccgcctg gtatatgatg gcatccggga
     2821 catcaggaaa gcagtgctga tgataaggac ccctgaggag ttggatgact ctgactttga
     2881 gacagaagat tttgatgtca gaagcaggac gagcgtccag acagaagacg atcagctgat
     2941 agctggccag agtgcccggg cgatcatggc tcagcttccc caggagcaaa aagcgaagat
     3001 tgcggaacag gtggccagct tccaggaaga aaagagcaag ctggatgctg aagtgtccaa
     3061 atgggacgac agtggcaatg acatcattgt gctggccaag cagatgtgca tgattatgat
     3121 ggagatgaca gactttaccc gaggtaaagg accactcaaa aatacatcgg atgtcatcag
     3181 tgctgccaag aaaattgctg aggcaggatc caggatggac aagcttggcc gcaccattgc
     3241 agaccattgc cccgactcgg cttgcaagca ggacctgctg gcctacctgc aacgcatcgc
     3301 cctctactgc caccagctga acatctgcag caaggtcaag gccgaggtgc agaatctcgg
     3361 cggggagctt gttgtctctg gggtggacag cgccatgtcc ctgatccagg cagccaagaa
     3421 cttgatgaat gctgtggtgc agacagtgaa ggcatcctac gtcgcctcta ccaaatacca
     3481 aaagtcacag ggtatggctt ccctcaacct tcctgctgtg tcatggaaga tgaaggcacc
     3541 agagaaaaag ccattggtga agagagagaa acaggatgag acacagacca agattaaacg
     3601 ggcatctcag aagaagcacg tgaacccggt gcaggccctc agcgagttca aagctatgga
     3661 cagcatctaa actcgagcgg ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg
     3721 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3781 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3841 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa
     3901 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4321 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4621 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5881 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5941 cacaccgcat aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
     6001 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     6061 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     6121 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     6181 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     6241 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     6301 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     6361 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     6421 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     6481 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     6541 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     6601 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     6661 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     6721 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     6781 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     6841 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     6901 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     6961 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     7021 cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7081 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
     7141 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7201 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc
     7261 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     7321 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     7381 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
     7441 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
     7501 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc
     7561 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
     7621 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
     7681 tggaatta
//
