LOCUS       sequence-328272-        6841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a caspase of Orbicella faveolata, which has higher sequence 
            similarity with human caspase-3 but has CARD domain and function as 
            an initiator caspase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6841)
  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 6841)
  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: OfCasp3a
FEATURES             Location/Qualifiers
     source          1..6841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     573..592
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(612..689)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     895..914
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     998..1017
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        998..1016
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1017..1041
                     /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             1056..1078
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2268..2285
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2339..2357)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2353..2400
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2575..2580,2581..2597,2598..2603))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2630..2648)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2716..2820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2821..2889,2890..3681)
                     /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(3039..3058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3852..4440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4341..4360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4594..4611
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4626..4765
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4691..4710)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4851..4873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4867..5058)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6370..6841,1..611))
                     /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 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
       61 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
      121 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
      181 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
      241 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
      301 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
      361 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
      421 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
      481 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
      541 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
      601 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
      661 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
      721 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
      781 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
      841 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
      901 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
      961 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     1021 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     1081 tacatatgga acagtctgat cgtgacattc tgcgtaaaaa ccgccaagat ctgctgaaag
     1141 acatggaagc gaaacgtgtg gcctctcgcc tgtacagtcg tggtattttt agtgaagaag
     1201 ataaagacga agttaactcc aaatcaacca cgaatgaaca gggtgaatgc ctgctggata
     1261 tcctgccgaa acgcggcccg aaagcattcc gtgctttttg tgatgttctg catgaactga
     1321 gtccgcacct ggaaagcctg ctgcgtccgg tccaggaagc gggtattccg agtgaaggca
     1381 ccgatggcgg taacaaaaat ccgatcagct ctacgtccaa tctgggtgca tcggaaagcg
     1441 ctgacgaagc ggatgccaaa ctgttcggct ttggcggtgg ctctgcaagt gctaaaccga
     1501 gttcctcaac cctggataaa gaaatcatct acaaaatgga ccgtagcacg cgcggcatcg
     1561 cagtcatcat caacaacaaa aactttctgc gttcgagcgg tatggatcgt tacccgcgca
     1621 atggcaccga tgtggaccgc gattccctgg ttaaactgtt ccgtatgctg aaattcgaag
     1681 ttaaaatcta caacgatcgc accaaagccg aaattcgtac catcacgaaa gaaatggcga
     1741 cgctgaacca ttcaaactac gatgccttca tttttagcat cctgacccac ggtgaagaag
     1801 gcgtgatcta cggtaccgat ggcacgattt ctatccgcga cctgaccgca gaattcaaat
     1861 attccacgtc actggctggt aaaccgaaac tgtttttctt tcaggcatgc caaggtcatg
     1921 aatatatgga cggcatggat gttaccgacg ctccgcagga aaataaagtg agcgttccgg
     1981 cggaagccga ttttctgtat gcctactcta ccgtcccggg ttattactcc tggcgtaact
     2041 cagtgaatgg ctcgtggttc attcaaagcc tgaccaaagt gtttgaagat aacgcggaac
     2101 gcatggacat cctgcgcatg ctgacgcgtg tcaatgccat ggttagcacc tacaaatctc
     2161 gcacgggcga ttattactcg gacagcaaac gtcaggtttc tagtattgtc tcaatgctgc
     2221 gcaaagaact gtatttcttt ccggaaaacg tggaacaatc gctggaacat caccatcacc
     2281 atcactgagg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     2341 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2401 ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca
     2461 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
     2521 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     2581 gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata
     2641 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2701 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2761 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2821 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2881 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2941 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3001 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3061 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3121 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3181 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3241 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3301 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3361 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3421 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3481 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3541 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3601 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3661 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3721 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3781 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3841 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3901 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3961 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4021 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4081 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4141 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4201 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4261 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4321 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4381 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4441 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4501 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4561 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4621 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     4681 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4741 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4801 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4861 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     4921 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     4981 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5041 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     5101 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     5161 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5221 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5281 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5341 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5401 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     5461 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     5521 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     5581 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     5641 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     5701 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     5761 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     5821 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     5881 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     5941 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     6001 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     6061 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     6121 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     6181 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     6241 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     6301 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     6361 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     6481 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     6541 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     6601 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     6661 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     6721 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     6781 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     6841 a
//
