LOCUS       sequence-375388-        4468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4468)
  AUTHORS   Zhou Q, Snipas S, Orth K, Muzio M, Dixit VM, Salvesen GS
  TITLE     Target protease specificity of the viral serpin CrmA. Analysis of 
            five caspases.
  JOURNAL   J Biol Chem. 1997 Mar 21. 272(12):7797-800.
  PUBMED    9065443
REFERENCE   2  (bases 1 to 4468)
  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. 1997 Mar 21. 272(12):7797-800."
COMMENT     Sequence Label: pET23b-Casp6-His
FEATURES             Location/Qualifiers
     source          1..4468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..99
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        80..98
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1048..1065
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1118..1136)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1132..1179
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1216..1671
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1303..1322)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1513..1534
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1697..1801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1802..1870,1871..2662)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2020..2039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2833..3421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3322..3341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3575..3592
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3607..3746
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3672..3691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3832..3854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3848..4039)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
       61 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      121 agaaataatt ttgtttaact ttaagaagga gatatacata tggctagctc ggcctcgggg
      181 ctccgcaggg ggcacccggc aggtggggaa gaaaacatga cagaaacaga tgccttctat
      241 aaaagagaaa tgtttgatcc ggcagaaaag tacaaaatgg accacaggag gagaggaatt
      301 gctttaatct tcaatcatga gaggttcttt tggcacttaa cactgccaga aaggcggggc
      361 acctgcgcag atagagacaa tcttacccgc aggttttcag atctaggatt tgaagtgaaa
      421 tgctttaatg atcttaaagc agaagaacta ctgctcaaaa ttcatgaggt gtcaactgtt
      481 agccacgcag atgccgattg ctttgtgtgt gtcttcctga gccatggcga aggcaatcac
      541 atttatgcat atgatgctaa aatcgaaatt cagacattaa ctggcttgtt caaaggagac
      601 aagtgtcaca gcctggttgg aaaacccaag atatttatca ttcaggcatg tcggggaaac
      661 cagcacgatg tgccagtcat tcctttggat gtagtagata atcagacaga gaagttggac
      721 accaacataa ctgaggtgga tgcagcctcc gtttacacgc tgcctgctgg agctgacttc
      781 ctcatgtgtt actctgttgc agaaggatat tattctcacc gggaaactgt gaacggctca
      841 tggtacattc aagatttgtg tgagatgttg ggaaaatatg gctcctcctt agagttcaca
      901 gaactcctca cactggtgaa caggaaagtt tctcagcgcc gagtggactt ttgcaaagac
      961 ccaagtgcaa ttggaaagaa gcaggttccc tgttttgcct caatgctaac taaaaagctg
     1021 catttctttc caaaatctaa tctcgagcac caccaccacc accactgaga tccggctgct
     1081 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1141 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1201 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1261 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1321 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1381 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1441 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1501 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1561 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1621 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2041 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2341 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2401 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2521 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3481 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3541 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3601 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     3661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3721 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3781 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3841 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3901 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3961 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4021 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4081 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4141 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4201 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4261 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4321 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4381 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4441 cagtaaggca accccgccag cctagccg
//
