LOCUS       sequence-248964-        6666 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 6666)
  AUTHORS   Martins LM, Iaccarino I, Tenev T, Gschmeissner S, Totty NF, Lemoine 
            NR, Savopoulos J, Gray CW, Creasy CL, Dingwall C, Downward J
  TITLE     The serine protease Omi/HtrA2 regulates apoptosis by binding XIAP 
            through a reaper-like motif.
  JOURNAL   J Biol Chem. 2002 Jan 4. 277(1):439-44.
  PUBMED    11602612
REFERENCE   2  (bases 1 to 6666)
  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. 2002 Jan 4. 277(1):439-44."
COMMENT     Sequence Label: pcDNA3-delta33HtrA-EGFP S306A
FEATURES             Location/Qualifiers
     source          1..6666
                     /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      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1985..2003)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1986..2003)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2023..2040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2029..2253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2299..2727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2386..2405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2596..2617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2736..2756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2741..3070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2921..3056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2983..3002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3137..3931
                     /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(3191..3210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3801..3820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4105..4226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4142..4161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4196..4215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4275..4291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4275..4291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4288..4310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4299..4315
                     /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(4323..4329,4330..4347,4348..4353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4368..4389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4506..4523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4677..5265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4757..4776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5436..6227,6228..6296))
                     /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     6059..6078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6297..6401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6476..6495)
                     /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 ggtacccgcc gccaccatgg acctccgggc cctgctgacg tcaggaactt ctgacccccg
      721 ggcccgagtg acttatggga cccccagtct ctgggcccgg ttgtctgttg gggtcactga
      781 accccgagca tgcctgacgt ctgggacccc gggtccccgg gcacaactga ctgcggtgac
      841 cccagatacc aggacccggg aggcctcaga gaactctgga acccgttcgc gcgcgtggct
      901 ggcggtggcg ctgggcgctg ggggggcagt gctgttgttg ttgtggggcg ggggtcgggg
      961 tcctccggcc gtcctcgccg ccgtccctag cccgccgccc gcttctcccc ggagtcagta
     1021 caacttcatc gcagatgtgg tggagaagac agcacctgcc gtggtctata tcgagatcct
     1081 ggaccggcac cctttcttgg gccgcgaggt ccctatctcg aacggctcag gattcgtggt
     1141 ggctgccgat gggctcattg tcaccaacgc ccatgtggtg gctgatcggc gcagagtccg
     1201 tgtgagactg ctaagcggcg acacgtatga ggccgtggtc acagctgtgg atcccgtggc
     1261 agacatcgca acgctgagga ttcagactaa ggagcctctc cccacgctgc ctctgggacg
     1321 ctcagctgat gtccggcaag gggagtttgt tgttgccatg ggaagtccct ttgcactgca
     1381 gaacacgatc acatccggca ttgttagctc tgctcagcgt ccagccagag acctgggact
     1441 cccccaaacc aatgtggaat acattcaaac tgatgcagct attgattttg gaaacgctgg
     1501 aggtcccctg gttaacctgg atggggaggt gattggagtg aacaccatga aggtcacagc
     1561 tggaatctcc tttgccatcc cttctgatcg tcttcgagag tttctgcatc gtggggaaaa
     1621 gaagaattcc tcctccggaa tcagtgggtc ccagcggcgc tacattgggg tgatgatgct
     1681 gaccctgagt cccagcatcc ttgctgaact acagcttcga gaaccaagct ttcccgatgt
     1741 tcagcatggt gtactcatcc ataaagtcat cctgggctcc cctgcacacc gggctggtct
     1801 gcggcctggt gatgtgattt tggccattgg ggagcagatg gtacaaaatg ctgaagatgt
     1861 ttatgaagct gttcgaaccc aatcccagtt ggcagtgcag atccggcggg gacgagaaac
     1921 actgacctta tatgtgaccc ctgaggtcac agaatgactc gagcatgcat ctagagggcc
     1981 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     2041 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2101 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2161 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2221 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2281 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2341 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2401 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2461 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2521 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2581 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2641 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2701 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2761 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2821 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2881 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2941 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3001 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3061 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3301 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3661 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4081 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4141 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4201 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4261 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4321 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4381 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4441 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4501 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4561 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4621 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4681 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4741 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4801 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4861 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4921 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4981 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5041 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5101 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5161 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5221 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5281 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5341 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5401 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5461 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5521 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5581 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5641 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5701 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5761 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5821 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5881 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5941 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6001 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6061 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6121 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6181 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6241 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6301 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6361 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6421 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6481 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6541 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     6601 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     6661 cgcgtt
//
