LOCUS       sequence-299764-        6750 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 6750)
  TITLE     Salvesen Casp1, Casp2 plasmids
REFERENCE   2  (bases 1 to 6750)
  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"
COMMENT     Sequence Label: pcDNA3-Casp2-Flag
FEATURES             Location/Qualifiers
     source          1..6750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      710..719
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2027..2050
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2069..2087)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2070..2087)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2107..2124)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2113..2337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2383..2811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2820..2840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2825..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3067..3086
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3221..4015
                     /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(3275..3294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3885..3904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4189..4310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4226..4245)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4280..4299
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4359..4375)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4359..4375)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4372..4394)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4383..4399
                     /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(4407..4413,4414..4431,4432..4437))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4452..4473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4590..4607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4761..5349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4841..4860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5520..6311,6312..6380))
                     /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     6143..6162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6381..6485)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6560..6579)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccaccatggc
      721 cgctgacagg ggacgcagga tattgggagt gtgtggcatg catcctcatc atcaggaaac
      781 tctaaaaaag aaccgagtgg tgctagccaa acagctgttg ttgagcgaat tgttagaaca
      841 tcttctggag aaggacatca tcaccttgga aatgagggag ctcatccagg ccaaagtggg
      901 cagtttcagc cagaatgtgg aactcctcaa cttgctgcct aagaggggtc cccaagcttt
      961 tgatgccttc tgtgaagcac tgagggagac caagcaaggc cacctggagg atatgttgct
     1021 caccaccctt tctgggcttc agcatgtact cccaccgttg agctgtgact acgacttgag
     1081 tctccctttt ccggtgtgtg agtcctgtcc cctttacaag aagctccgcc tgtcgacaga
     1141 tactgtggaa cactccctag acaataaaga tggtcctgtc tgccttcagg tgaagccttg
     1201 cactcctgaa ttttatcaaa cacacttcca gctggcatat aggttgcagt ctcggcctcg
     1261 tggcctagca ctggtgttga gcaatgtgca cttcactgga gagaaagaac tggaatttcg
     1321 ctctggaggg gatgtggacc acagtactct agtcaccctc ttcaagcttt tgggctatga
     1381 cgtccatgtt ctatgtgacc agactgcaca ggaaatgcaa gagaaactgc agaattttgc
     1441 acagttacct gcacaccgag tcacggactc ctgcatcgtg gcactcctct cgcatggtgt
     1501 ggagggcgcc atctatggtg tggatgggaa actgctccag ctccaagagg tttttcagct
     1561 ctttgacaac gccaactgcc caagcctaca gaacaaacca aaaatgttct tcatccaggc
     1621 ctgccgtgga gatgagactg atcgtggggt tgaccaacaa gatggaaaga accacgcagg
     1681 atcccctggg tgcgaggaga gtgatgccgg taaagaaaag ttgccgaaga tgagactgcc
     1741 cacgcgctca gacatgatat gcggctatgc ctgcctcaaa gggactgccg ccatgcggaa
     1801 caccaaacga ggttcctggt acatcgaggc tcttgctcaa gtgttttctg agcgggcttg
     1861 tgatatgcac gtggccgaca tgctggttaa ggtgaacgca cttatcaagg atcgggaagg
     1921 ttatgctcct ggcacagaat tccaccggtg caaggagatg tctgaatact gcagcactct
     1981 gtgccgccac ctctacctgt tcccaggaca ccctcccaca ctcgaggact acaaggacga
     2041 cgatgacaag tgatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     2101 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2161 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2221 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2281 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2341 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2821 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3181 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3241 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3301 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3361 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3421 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3481 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3541 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3601 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3661 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3721 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3781 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3841 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3901 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3961 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4021 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4081 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4141 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4201 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4261 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4321 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4381 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4441 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4501 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4561 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5161 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6121 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6481 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6541 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6601 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6661 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     6721 ttcgcgatgt acgggccaga tatacgcgtt
//
