LOCUS       sequence-368768-        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the HMP1 F-actin Binding domain (677-927) 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 5723)
  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 5723)
  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-HMP1_FABD
FEATURES             Location/Qualifiers
     source          1..5723
                     /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(1773..1795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1940..1958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2026..2130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2131..2199,2200..2991)
                     /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(2349..2368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3162..3750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3651..3670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3904..3921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3994..4071
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4072..5154
                     /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(4091..4110)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5203..5208,5209..5226,5227..5233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5241..5257
                     /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     5265..5281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5265..5281
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5306..5328)
                     /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 aatctgatga
      961 gacgattgcc agaagaagag aaaaagaaaa ttcaagcaca aattgatatc ttcaaagtaa
     1021 cacaaacacg attcgaaaga gaagttgcca agtgggatga aacaggaaat gatattatat
     1081 cattggccaa taatatgtgt aagattatga tgagtatgac agaattcaca agaggttgtg
     1141 ggccattgaa aacgacaatg gatgttatcc gagcagcaca agaaatctca cttaatggat
     1201 caaaactaaa tgcattggca cggcagattg gagaagaaag tgctgattcg cagacgaaaa
     1261 aggatttgct cgcatatttg tcacaaatca ctctatattg tcagcaactg aatatttgta
     1321 gtaaggtgaa agctgatgtg actcaagttg gaaatgagct tgtcgtatct gcacttgact
     1381 ctgcaatgtc tcttattcaa acggctcgaa atcttttaac tgctgttgta cagactgtta
     1441 aagctgcata cattgccagt accaagttcc gtcgtcctaa tgccaacagt gtacgtgtcg
     1501 agtggcgaat ggctccacca aagaaacaac cgctaattcg accacaaaag aataatgcta
     1561 taatcagacg ggcatccgaa cggcgacctt tgcaaccagc taaagtgctc gccgagttta
     1621 cgcgtaacga gattgagacg ggtcgtgata gtgacgacga ggaacttgat cgccggcatc
     1681 aacaaagaat aaacggtcgt ttatgaactc gagcggccgc atcgtgactg actgacgatc
     1741 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1801 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1861 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     1921 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1981 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2041 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2101 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2161 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2221 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2281 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2341 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2401 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2461 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2521 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2581 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2641 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2701 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2761 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2821 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2881 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2941 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3001 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3301 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3781 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3841 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3901 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3961 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
     4021 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     4081 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     4141 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     4201 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     4261 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     4321 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     4381 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     4441 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     4501 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     4561 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     4621 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     4681 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     4741 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     4801 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     4861 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     4921 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     4981 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     5041 gtgaaaagaa aaaccacgct agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5101 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5161 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     5221 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     5281 ccatgattac ggattcactg gccatcgttt tacaacgtcg tgactgggaa aaccctggcg
     5341 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5401 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     5461 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     5521 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     5581 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     5641 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     5701 ttatttttga tggcgttgga att
//
