LOCUS       sequence-47470-e        8884 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 8884)
  AUTHORS   Miedel MT, Graf NJ, Stephen KE, Long OS, Pak SC, Perlmutter DH, 
            Silverman GA, Luke CJ
  TITLE     A pro-cathepsin L mutant is a luminal substrate for 
            endoplasmic-reticulum-associated degradation in C. elegans.
  JOURNAL   PLoS One. 2012;7(7):e40145. Epub 2012 Jul 2.
  PUBMED    22768338
REFERENCE   2  (bases 1 to 8884)
  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: "PLoS One.";
            date: "2012"; volume: "7(7)"; pages: "e40145. Epub 2012 Jul 2"
COMMENT     Sequence Label: pKS2301
FEATURES             Location/Qualifiers
     source          1..8884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4792..4820)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5568..5588
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6458..6480)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6580..6599
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6677..6695)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6763..6867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6868..6936,6937..7728)
                     /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     complement(7086..7105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7899..8487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8388..8407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8641..8658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8775..8796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8811..8816,8817..8834,8835..8841)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8849..8865
                     /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)."
     primer_bind     8854..8876
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8873..8884,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8873..8884,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttagctttg tggcgaccac cacgtgtcta ctgataaatt
       61 aacagattgt actactttgc atgtaatgtg ctagaggacg gttctcttat catcacaggc
      121 actgacgaca ctgccaaagt tttacaagct aatttattag cgctcttcgg ccgcaatggc
      181 acgatttgaa ttggtttatt gattccagag actagtttca gtttttgata agaatgaatt
      241 attattcatt ttctagttta tgaatatata tgtaactagg gaagcctatt tatgattttt
      301 ctaatttttt aaaattgtta ttttagagag gtgtgaaatt aaaatgatat atattttcga
      361 gtactttaaa aaataaaaac ggtacatcct atattaaatt tttgatttta gatccttcca
      421 agttattgac aagtattaac tgcacttctc ataaaaattt ttcaaataga tctatttagt
      481 ctggaatagg aatacttttg gcaattttct tgagaatcat agaagacgaa tttctgaact
      541 gctctttgtg aagccacgcc catatattgg catttttgta tccaattttt tgtagagtac
      601 ctgaatcatt atcactacat ctgctaaaca atatatggaa gttctatgag aggtgaaagg
      661 taaatcatat aacaactgta ataacataga aatgattgta gtatatttcc aattaggctg
      721 agttgatctt attgaagagt ttgaatcata gcaatttttt cctcttttag tatttttcaa
      781 aacaaatttt cacaatgtta gatgtagtat ttagcattaa ggcatctgta atctcccaga
      841 aatgttttgg gaataaaacc ctttgactct ttggtccttt tttgctccaa ataatatctg
      901 attttaaccc gttctgatct actttttacc tttcgatcgt cacatccgcg tacacgtttc
      961 tcttgctctc aattgcttat ctttatggcc ttttccccca ctttttgtcg actatcatgc
     1021 ggacgtggct tccttgctcc ctcccggttt ttttgaaaac acttgtgaat ttattttttg
     1081 tggtcaaaat tacgaaacat ttgaattctc aatctattta tagaccgggt gtggtatgtt
     1141 atgccatgtt tttgtcattg ccaaatgaac atcgatattt taattttctg aacttaccgt
     1201 accccctgcg ttactatttt atttttaatt tgtataattt cattgctata atttgttgtc
     1261 cttgaaacat tgaagatatc gaagaaataa tgacaaaaat cagatagaac aatttttcct
     1321 actcatcaac tcacattttt ctattttctc ttgtgcgaaa aaaatagtat attgggaata
     1381 tttatagtat tgcaactttt ctcttagttc gcagagttgc aatactacta gatccctaat
     1441 agaggaaata tggtgaccag ctttttgcaa ttggaaagtt gggaaaaaag tatatatatt
     1501 aatagactta cactcttgta gttcgtagaa tttttcagga aatctgacac gcaagacttt
     1561 ttcaaaaagt tttttttttt aatttggcaa tctgccaaaa cgtttactga acattttgaa
     1621 aacatctacc agtaccttta acacaacgaa aaattgttct aagcaatatt aactgtgata
     1681 attgatgtta agacctgaaa tttaatcaca aatttcacct ggaaatgaca gtaatcggct
     1741 tctaaatact gtcgacacaa gtgccatacc taagattcca atcctattac catgtttccc
     1801 aatttcattc tctttcgtat ctctatttct cttctttatt atcaattact ttttatcagt
     1861 tcttcgtttg ctttaacatc aagagcacat agcgctctct cgtctctctc tcatgctctt
     1921 tttgaacttt tacaaaaaac tatttccggt tgttttgatt cttggaattt gaaataattt
     1981 tcagtgatta aatcatgtct ctgaggcctt gggtgctcgt cgttggactt ttattgattt
     2041 ctccgtctag aggatccccg ggattggcca aaggacccaa aggtatgttt cgaatgatac
     2101 taacataaca tagaacattt tcaggaggac ccttggctag cagaatgaac cgattcattc
     2161 ttctggcact ggttgccgcc gtcgtcgccg tcaattcggc caagctgtcc cgtcaaatcg
     2221 agtcggccat cgagaaatgg gacgactata aggaggactt tgataaggag tactcggaga
     2281 gcgaggagca gacctatatg gaggcatttg tcaagaatat gatccatatt gagaatcaca
     2341 acagagatca ccgactcgga agaaagacat tcgagatggg attgaatcat attgctgact
     2401 tggtaagaat atgcatttca atgcaccatg agctgcggcg tccgacgttt tgagctccgg
     2461 ggactcaaat tctggctccg aggaccgaag tctctagctc cagggtccca agttttgagc
     2521 tgcggggttt caaaattcaa gctccggttc ttaagtaacg agctccggga tcacaatttt
     2581 tgagcccaag gatctcaagt ttggagctcc ggagtcccaa gttcctaaag cgaggatttt
     2641 caagttttcc ttttttgcat ttcctaattt cgacagataa gaaaataaat accccctata
     2701 aaaacttcaa aaagctgttc tctgtctcgt tccactatca aaatctcacc cagcgagaaa
     2761 aaaaaagata aagatatgaa atcctcgtct cgtgactgac caacagctgg cttatcacac
     2821 caccacactg acaaaacaca cgaaggtcat ttgattttca aactcttgtt tttttcagcc
     2881 attcagccaa taccgcaaac tcaacggtta cagacgtctc ttcggtgact ccagaatcaa
     2941 gaactcctcc tctttcttgg ctccattcaa tgttcaggta attttttgag aggggttttg
     3001 ttggggtcca gacactaact ttctcgtcga tcaaaactat aaaactattt gcataaacaa
     3061 aaacgttttc aacgaccgat aaaaataatt cttcgctgac cgtatttttt gaattatcat
     3121 ctctgatgat tgatacgctt gaaaattgaa tttcgcaatg ttttggtcac cgaaatgttt
     3181 aaaaacaaga ttcaaatggt tagaactgtt tttgctgaaa tttgaacacc agagcttaga
     3241 gctgtgatga acttctaggg gataagccta agcctgagct tcagtctaag cctaaatcta
     3301 agcctacgtc taagtgtacg cctaagctta agcctcagcc taagcctcag cctaagccta
     3361 agcctaggct tgaaactgat tgtgagttga tggatttggc tcagtccagg aagaagtttg
     3421 gtctatgcct aagccttagc ataagcttga gcttcccaag ctttcggcta aaccaaaccc
     3481 tcagcttaaa ctcaagccta aacctattca attttcaggt cccagatgag gttgactggc
     3541 gtgataccca cctcgtcact gatgtcaaga accaaggaat gtgcggatcg tgctgggcct
     3601 tctccgccac cggagccctc gaaggacaac acgctcgcaa gctgggacaa ctcgtctccc
     3661 tttccgagca aaacctcgtc gactgctcta ccaagtacgg aaaccacgga tgcaacggag
     3721 gactcatgga tcaagctttc gagtacattc gtgacaacca tggtgtcgac accgaggagt
     3781 catacccata caagggacgt gacatgaagt gtcacttcaa caagaagacc gtcggagctg
     3841 atgataaggg atacgttgac accccagaag gagatgagga gcaacttaag atcgctgtcg
     3901 ccacccaagg accaatctct attgctatcg acgccggaca ccgcagcttc caactttaca
     3961 agaagggtat gttttgtggg aagatgtgag gggggggggg gtactagttt ttattgacgg
     4021 gtcagtttta tttgcatttt ggatgataat gataagaaag ctagttactc gggtttttca
     4081 tcagagacag agagggcaat agtgataaga gaactcgtgt ttttgaatga taccctcttg
     4141 acaataatag tttgggttcg aacttgaatg tgaaagtggt gaaggaattt ataaattgtt
     4201 tagtagcaaa gtataacccc atcagcctag gccaaagtct aagcttaagc ttacgattaa
     4261 gcctaagcct aagcctaagc ttacgcctaa gcctaagcct acgcctaaga ctaagcctaa
     4321 acctaagcga aagcctaagc ctaagcctga gcctaagctt aagcctaagc ctgagtctaa
     4381 gccgtagctc aagactaagc ctgagtctaa tcccaagcct agctcataag cctaagctta
     4441 aacaaatgcc taaaccaaag cctaagccta ggcctaaaac taggcctaca gcataaactc
     4501 agcctaagtt ccaccgaata tcatccattt tcaggagtct actacgatga ggaatgctca
     4561 tccgaagagc tcgaccacgg agtgcttctc gtcggatacg gaaccgaccc agagcacgga
     4621 gactactgga ttgtcaagaa ctcgtgggga gctggatggg gagagaaggg atacatccgt
     4681 atcgcccgta accgcaacaa tcactgcgga gtcgccacca aggccagtta tccattggtc
     4741 ggaggaggag gaagcgctag catgagtaaa ggagaagaac ttttcactgg agttgtccca
     4801 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     4861 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     4921 cctgttccat gggtaagttt aaacatatat atactaacta accctgatta tttaaatttt
     4981 cagccaacac ttgtcactac tttcggttat ggtcttcaat gcttcgccag atacccagat
     5041 catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
     5101 actatatttt tcaaagatga cgggaactac aagacacgta agtttaaaca gttcggtact
     5161 aactaaccat acatatttaa attttcaggt gctgaagtca agtttgaagg tgataccctt
     5221 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
     5281 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
     5341 ggaatcaaag ttgtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     5401 cagaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     5461 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     5521 taccaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     5581 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata ggagctccgc
     5641 atcggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt
     5701 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     5761 atcaaaaaaa cttcttctta atttctttgt tttttagctt cttttaagtc acctctaaca
     5821 atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt
     5881 gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac
     5941 ttcttatgtt ttttttactt ctgataaatt ttttttgaaa catcatagaa aaaaccgcac
     6001 acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca
     6061 cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt
     6121 ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg
     6181 gtttccccta ttgtttgtca agagtttcga ggacggcgtt tttcttgcta aaatcacaag
     6241 tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa
     6301 ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat
     6361 gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc
     6421 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6481 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6541 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     6601 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6661 tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6721 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6781 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6841 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6901 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6961 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     7021 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     7081 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     7141 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     7201 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7261 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     7321 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7381 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7441 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7501 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7561 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7621 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7681 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7741 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8041 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8281 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8581 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8641 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8701 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8761 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8821 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8881 agct
//
