LOCUS       sequence-328270-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a caspase of Porites astreoides, which has higher sequence 
            similarity with human caspase-3 but function similar to caspase-6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  AUTHORS   Shrestha S, Tung J, Grinshpon RD, Swartz P, Hamilton PT, Dimos B, 
            Mydlarz L, Clark AC
  TITLE     Caspases from scleractinian coral show unique regulatory features.
  JOURNAL   J Biol Chem. 2020 Oct 23;295(43):14578-14591. doi: 
            10.1074/jbc.RA120.014345. Epub 2020 Aug 11.
  PUBMED    32788218
REFERENCE   2  (bases 1 to 6559)
  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.RA120.014345"; journalName: "J Biol Chem"; date: 
            "2020-10-23- 23"; volume: "295"; issue: "43"; pages: "14578-14591"
COMMENT     Sequence Label: PaCasp3
FEATURES             Location/Qualifiers
     source          1..6559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     917..936
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(956..1033)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1239..1258
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1342..1361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1342..1360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1361..1385
                     /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)."
     RBS             1400..1422
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2330..2347
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2401..2419)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2415..2462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2637..2642,2643..2659,2660..2665))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2692..2710)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2778..2882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2883..2951,2952..3743)
                     /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(3101..3120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3914..4502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4403..4422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4656..4673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4688..4827
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4753..4772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4913..4935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4929..5120)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6432..6559,1..955))
                     /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"
ORIGIN
        1 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
       61 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
      121 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
      181 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
      241 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
      301 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
      361 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
      421 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
      481 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
      541 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
      601 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
      661 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
      721 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
      781 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
      841 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
      901 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
      961 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     1021 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     1081 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     1141 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     1201 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     1261 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     1321 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     1381 attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgtcaaaatt
     1441 taacaccgtg gtttcgggct gcgcgggtaa tattgtcgtg ggcgaccatg ccgttctgaa
     1501 cgttggtgcc agctctaccg atgcacgtcc gcgtaaacag ccgccgtcag aagaagttcc
     1561 gccgaccggt ccgagccagg tccactgtca agaatctgca ggcgatggta tgaactacat
     1621 cgctaaatct ggttacgttc tggtcatcaa caattacctg tttccgcagc gtctgaacgt
     1681 ggaacgcacc ggctcaaatg atgacgttaa aaacctgacg aacctgtttg atgactttgg
     1741 tttccgttcg cgcgtgcagg ataaccaaac ccgttcagaa atgctgtcgc tgctgaaaga
     1801 tacggcagaa aaagacttca gtaaatacga ttgcttcgtg tgtgttattc tgagccatgg
     1861 ctctaaagac ggcatctacg gtaccgatga cgaagttatt aatatcgaag cgattacgag
     1921 tctgtttcgt cgcgatgaat gcccgtccct ggaaggcaaa ccgaaaattt tcctgatcca
     1981 ggcctgtcgt ggtaaccaac gcgatattgt cccggtggaa tcagattcgg acccgatggt
     2041 gtttagtaat ccgtccctgc cggcggatgc cgactttctg atctgcttcg caagtgctcc
     2101 gggccaccag tcctatcgcc aaccgctgct gggtagctgg ttcatttctg cagtgtttga
     2161 cgttttcaaa gaacatgctg aacgtgaaca catcatggat atgatgctgc gcgtcaacaa
     2221 tcaggtggcc ggctattaca gccgtgatgg tctgaaacag atgccgtgcc aaatctgtat
     2281 gctgaccaag aaagtgtttt tccagccgcg ctacggtagt tccctggaac atcaccatca
     2341 ccatcactga ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     2401 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2461 tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac
     2521 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     2581 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     2641 aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga
     2701 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2761 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2881 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2941 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3001 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3061 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3121 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3421 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4021 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4621 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4681 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     4741 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     4801 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4861 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4921 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     4981 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     5041 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     5101 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     5161 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     5221 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     5281 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     5341 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     5401 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     5461 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     5521 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     5581 acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     5641 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     5701 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc
     5761 aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata
     5821 gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg
     5881 ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt
     5941 gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca
     6001 tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg
     6061 cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct
     6121 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     6181 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     6241 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     6301 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     6361 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     6421 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6481 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     6541 ttcaccagtg agacgggca
//
