LOCUS       sequence-328273-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a caspase of Orbicella faveolata, 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 6562)
  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 6562)
  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: OfCasp3b
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     562..581
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(601..678)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     884..903
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     987..1006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        987..1005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1006..1030
                     /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             1045..1067
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1978..1995
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2049..2067)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2063..2110
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2285..2290,2291..2307,2308..2313))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2340..2358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2426..2530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2531..2599,2600..3391)
                     /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(2749..2768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3562..4150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4051..4070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4304..4321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4336..4475
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4401..4420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4561..4583
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4577..4768)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6080..6562,1..600))
                     /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 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
       61 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
      121 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
      181 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
      241 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
      301 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
      361 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
      421 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
      481 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
      541 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
      601 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
      661 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      721 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      781 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      841 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      901 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      961 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     1021 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat acatatgagc
     1081 tctaaattta aaattcatgt ggcagattgc agtggcgctg tggttatcgg tgaccacgcg
     1141 aacctgacgg tggaagcacc ggttgccggt gcgggtccgc agcgtcgccc ggttcgtgaa
     1201 caagccgcag cagctaccac gaccagtacc catgcaccgt gtcaagaaag ttccggcgat
     1261 gacatcaact acaaagctaa atccggttac gtgctggtta tcaacaacta catctttccg
     1321 cgtcgcgatg acgtggaacg tacgggcagt aacgatgacg ttcgcaatct gaccagcctg
     1381 ttcgatgact ttaacttccg tcatgtcgtg gaagataatc agaccggtag ccaaatgatc
     1441 aaactgctgc agacgaccgc agaaaaagat tttagccgct atgactgctt cgtctgtgtg
     1501 attctgtctc acggcaagaa agatggcatc tacggtacgg acgaagaact gattaaaatc
     1561 gaagcaatta cctcactgtt tcgtcgcaac gaatgcccgt cgctggaagg caaaccgaaa
     1621 atctttctgt tccaggcttg tcgtggttca cgtcgcgata ccgttccggt cgaatcagat
     1681 tcggacccga ttccgctgtc atcgagctct ctgccggcag atgctgactt tctgatctgc
     1741 ttcgcgagcg ccccggatca tgaatcttat cgccaagccc acctgggttc ttggtttatt
     1801 agttccgttg tcgatgtgtt caaagaatac gcggaacgtg aacatctgat ggatatgatg
     1861 ctgcgcgtca acaatcacgt ggccggcttc tattcaaaag aaggtctgaa acagatcccg
     1921 tgccaagttt gtatgctgcg taagaaagtg tttttcgatc cgaaatactc gctggaacat
     1981 caccatcacc atcactgagg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2041 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2101 gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc
     2161 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     2221 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     2281 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     2341 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2521 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2581 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2641 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2701 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2761 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2821 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2881 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2941 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3001 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3061 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3121 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3181 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3241 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3301 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3361 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3421 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3481 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3541 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3601 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3661 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4021 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4081 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4141 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4201 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4261 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4321 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4381 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4441 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4501 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4561 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4621 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4681 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4741 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4801 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4861 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4921 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4981 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     5041 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     5101 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     5161 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     5221 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     5281 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     5341 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     5401 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     5461 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     5521 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     5581 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     5641 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     5701 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     5761 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5821 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5881 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5941 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     6001 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     6061 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6121 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     6181 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     6241 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     6301 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     6361 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     6421 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     6481 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     6541 tgcgagtgag atatttatgc ca
//
