LOCUS       sequence-424719-        6782 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 6782)
  AUTHORS   Plun-Favreau H, Klupsch K, Moisoi N, Gandhi S, Kjaer S, Frith D, 
            Harvey K, Deas E, Harvey RJ, McDonald N, Wood NW, Martins LM, 
            Downward J
  TITLE     The mitochondrial protease HtrA2 is regulated by Parkinson's 
            disease-associated kinase PINK1.
  JOURNAL   Nat Cell Biol. 2007 Nov;9(11):1243-52. doi: 10.1038/ncb1644. Epub 
            2007 Sep 30.
  PUBMED    17906618
REFERENCE   2  (bases 1 to 6782)
  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: "10.1038/ncb1644"; 
            journalName: "Nat Cell Biol"; date: "2007-11"; volume: "9"; issue: 
            "11"; pages: "1243-52"
COMMENT     Sequence Label: pcDNA3-HtrA2-FLAG
FEATURES             Location/Qualifiers
     source          1..6782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     243..262
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             397..1191
                     /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(451..470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1061..1080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1365..1486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1402..1421)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1456..1475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1535..1551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1535..1551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1548..1570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1559..1575
                     /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(1583..1589,1590..1607,1608..1613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1628..1649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1766..1783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1937..2525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2017..2036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2696..3487,3488..3556))
                     /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     3319..3338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3557..3661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3736..3755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3927..4306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4307..4510
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4461..4481
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4555..4574
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4555..4573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5973..5996
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6027..6045)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6028..6045)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6065..6082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6071..6295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6341..6769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6428..6447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6638..6659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(6778..6782,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
      361 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
      421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
      481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
      541 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
      601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
      661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
      721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
      781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
      841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
      901 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
      961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     1081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     1141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     1201 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     1261 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     1321 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     1381 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1441 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     1501 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1561 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1621 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1681 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1741 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1801 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1861 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1921 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1981 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2041 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2101 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2161 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2221 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2281 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2341 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2401 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2461 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2521 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2581 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2641 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2701 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2761 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2821 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2881 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2941 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3001 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3061 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3121 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3181 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3241 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3301 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3361 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3421 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3481 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3541 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3601 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3661 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     3721 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     3781 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     3841 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     3901 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     3961 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     4021 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     4081 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     4141 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     4201 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     4261 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     4321 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     4381 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     4441 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     4501 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     4561 gactcactat agggagaccc aagcttggta ccaccaccat ggctgcgccg agggcggggc
     4621 gaggtgcagg ctggagcctt cgggcatggc gggctttggg gggcattcgc tgggggagga
     4681 gaccccgttt gacccctgac ctccgggccc tgctgacgtc aggaacttct gacccccggg
     4741 cccgagtgac ttatgggacc cccagtctct gggcccggtt gtctgttggg gtcactgaac
     4801 cccgagcatg cctgacgtct gggaccccgg gtccccgggc acaactgact gcggtgaccc
     4861 cagataccag gacccgggag gcctcagaga actctggaac ccgttcgcgc gcgtggctgg
     4921 cggtggcgct gggcgctggg ggggcagtgc tgttgttgtt gtggggcggg ggtcggggtc
     4981 ctccggccgt cctcgccgcc gtccctagcc cgccgcccgc ttctccccgg agtcagtaca
     5041 acttcatcgc agatgtggtg gagaagacag cacctgccgt ggtctatatc gagatcctgg
     5101 accggcaccc tttcttgggc cgcgaggtcc ctatctcgaa cggctcagga ttcgtggtgg
     5161 ctgccgatgg gctcattgtc accaacgccc atgtggtggc tgatcggcgc agagtccgtg
     5221 tgagactgct aagcggcgac acgtatgagg ccgtggtcac agctgtggac cccgtggcag
     5281 acatcgcaac gctgaggatt cagactaagg agcctctccc cacgctgcct ctgggacgct
     5341 cagctgatgt ccggcaaggg gagtttgttg ttgccatggg aagtcccttt gcactgcaga
     5401 acacgatcac atccggcatt gttagctctg ctcagcgtcc agccagagac ctgggactcc
     5461 cccaaaccaa tgtggaatac attcaaactg atgcagctat tgattttgga aactctggag
     5521 gtcccctggt taacctggat ggggaggtga ttggagtgaa caccatgaag gtcacagctg
     5581 gaatctcctt tgccatccct tctgatcgtc ttcgagagtt tctgcatcgt ggggaaaaga
     5641 agaactcctc ctccggaatc agtgggtccc agcggcgcta cattggggtg atgatgctga
     5701 ccctgagtcc cagcatcctt gctgaactac agcttcgaga accaagcttt cccgatgttc
     5761 agcatggtgt actcatccat aaagtcatcc tgggctcccc tgcacaccgg gctggtctgc
     5821 ggcctggtga tgtgattttg gccattgggg agcagatggt acaaaatgct gaagatgttt
     5881 atgaagctgt tcgaacccaa tcccagttgg cagtgcagat ccggcgggga cgagaaacac
     5941 tgaccttata tgtgacccct gaggtcacag aagactacaa agacgatgac gacaaatgac
     6001 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     6061 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6121 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6181 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6241 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     6301 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     6361 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6421 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6481 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6541 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6601 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6661 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6721 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     6781 at
//
