LOCUS       sequence-220149-        6848 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 6848)
  AUTHORS   Stennicke HR, Salvesen GS
  TITLE     Biochemical characteristics of caspases-3, -6, -7, and -8.
  JOURNAL   J Biol Chem. 1997 Oct 10. 272(41):25719-23.
  PUBMED    9325297
REFERENCE   2  (bases 1 to 6848)
  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 Oct 10. 272(41):25719-23."
COMMENT     Sequence Label: pcDNA3-Casp8
FEATURES             Location/Qualifiers
     source          1..6848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2362..2385
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2403..2421)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2404..2421)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2441..2458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2447..2671
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2717..3145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2804..2823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3014..3035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3154..3174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3159..3488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3339..3474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3401..3420
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3555..4349
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3609..3628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4219..4238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4523..4644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4560..4579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4614..4633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4693..4709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4693..4709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4706..4728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4717..4733
                     /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)."
     promoter        complement(join(4741..4747,4748..4765,4766..4771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4786..4807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4924..4941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5095..5683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5175..5194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5854..6645,6646..6714))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6477..6496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6715..6819)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctaccgg
      901 taccatggac ttcagcagaa atctttatga tattggggaa caactggaca gtgaagatct
      961 ggcctccctc aagttcctga gcctggacta cattccgcaa aggaagcaag aacccatcaa
     1021 ggatgccttg atgttattcc agagactcca ggaaaagaga atgttggagg aaagcaatct
     1081 gtccttcctg aaggagctgc tcttccgaat taatagactg gatttgctga ttacctacct
     1141 aaacactaga aaggaggaga tggaaaggga acttcagaca ccaggcaggg ctcaaatttc
     1201 tgcctacagg gtcatgctct atcagatttc agaagaagtg agcagatcag aattgaggtc
     1261 ttttaagttt cttttgcaag aggaaatctc caaatgcaaa ctggatgatg acatgaacct
     1321 gctggatatt ttcatagaga tggagaagag ggtcatcctg ggagaaggaa agttggacat
     1381 cctgaaaaga gtctgtgccc aaatcaacaa gagcctgctg aagataatca acgactatga
     1441 agaattcagc aaagagagaa gcagcagcct tgaaggaagt cctgatgaat tttcaaatgg
     1501 ggaggagttg tgtggggtaa tgacaatctc ggactctcca agagaacagg atagtgaatc
     1561 acagactttg gacaaagttt accaaatgaa aagcaaacct cggggatact gtctgatcat
     1621 caacaatcac aattttgcaa aagcacggga gaaagtgccc aaacttcaca gcattaggga
     1681 caggaatgga acacacttgg atgcaggggc tttgaccacg acctttgaag agcttcattt
     1741 tgagatcaag ccccacgatg actgcacagt agagcaaatc tatgagattt tgaaaatcta
     1801 ccaactcatg gaccacagta acatggactg cttcatctgc tgtatcctct cccatggaga
     1861 caagggcatc atctatggca ctgatggaca ggaggccccc atctatgagc tgacatctca
     1921 gttcactggt ttgaagtgcc cttcccttgc tggaaaaccc aaagtgtttt ttattcaggc
     1981 ttgtcagggg gataactacc agaaaggtat acctgttgag actgattcag aggagcaacc
     2041 ctatttagaa atggatttat catcacctca aacgagatat atcccggatg aggctgactt
     2101 tctgctgggg atggccactg tgaataactg tgtttcctac cgaaaccctg cagagggaac
     2161 ctggtacatc cagtcacttt gccagagcct gagagagcga tgtcctcgag gcgatgatat
     2221 tctcaccatc ctgactgaag tgaactatga agtaagcaac aaggatgaca agaaaaacat
     2281 ggggaaacag atgcctcagc ctactttcac actaagaaaa aaacttgtct tcccttctga
     2341 ttaaggatcc aagcttctag agactacaaa gacgatgacg acaagtgact agagggccct
     2401 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     2461 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2521 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2581 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2641 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2701 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2761 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3061 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3121 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3721 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4081 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4381 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4441 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4501 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4561 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4621 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4681 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4741 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4801 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5521 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5701 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5761 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5821 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5881 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5941 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6001 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6061 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6121 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6181 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6241 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6301 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6361 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6421 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6481 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6541 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6601 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6661 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6721 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6781 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6841 acctgacg
//
