LOCUS       sequence-48349-e        6211 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 6211)
  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 6211)
  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: pKS2236
FEATURES             Location/Qualifiers
     source          1..6211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2119..2147)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2895..2915
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3785..3807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3907..3926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4004..4022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4090..4194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4195..4263,4264..5055)
                     /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(4413..4432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5226..5814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5715..5734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5968..5985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6102..6123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6138..6143,6144..6161,6162..6168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6192
                     /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     6181..6203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6200..6211,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6200..6211,1..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 cattcttgta 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 aatctagagg atccccggga ttggccaaag gacccaaagg tatgtttcga
     2041 atgatactaa cataacatag aacattttca ggaggaccct tggctagcat gagtaaagga
     2101 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     2161 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     2221 aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa catatatata
     2281 ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt cggttatggt
     2341 cttcaatgct tcgccagata cccagatcat atgaaacagc atgacttttt caagagtgcc
     2401 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     2461 acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt ttcaggtgct
     2521 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     2581 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     2641 tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa catgatttta
     2701 ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa cattgaagat
     2761 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     2821 cttttaccag acaaccatta cctgtcctac caatctgccc tttcgaaaga tcccaacgaa
     2881 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     2941 gatgaactat acaaatagga gctccgcatc ggccgctgtc atcagatcgc catctcgcgc
     3001 ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct
     3061 gtgctcccac cccctatttt tgttattatc aaaaaaactt cttcttaatt tctttgtttt
     3121 ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat
     3181 tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc
     3241 aaaatctaca caatgttctg tgtacacttc ttatgttttt tttacttctg ataaattttt
     3301 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     3361 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     3421 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     3481 taattttcct gcttttgctt tttgggggtt tcccctattg tttgtcaaga gtttcgagga
     3541 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     3601 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     3661 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     3721 tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     3781 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3841 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     3901 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     3961 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     4021 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4081 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4141 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4201 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4261 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4321 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4381 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4441 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4501 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5341 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5581 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     5641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5881 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5941 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6001 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6061 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6181 agcggataac aatttcacac aggaaacagc t
//
