LOCUS       sequence-47456-e        6371 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 6371)
  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 6371)
  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: pOL2339
FEATURES             Location/Qualifiers
     source          1..6371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2142..2366
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVFRLRG"
     CDS             2403..3113
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2548..2566)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2816..2835
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3945..3967)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4067..4086
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4164..4182)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4250..4354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4355..4423,4424..5215)
                     /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(4573..4592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5386..5974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5875..5894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6128..6145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6262..6283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6298..6303,6304..6321,6322..6328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6336..6352
                     /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     6341..6363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6360..6371,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6360..6371,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgca gctttgtggc gaccaccacg tgtctactga
       61 taaattaaca gattgtacta ctttgcatgt aatgtgctag aggacggttc tcttatcatc
      121 acaggcactg acgacactgc caaagtttta caagctaatt tattagcgct cttcggccgc
      181 aatggcacga tttgaattgg tttattgatt ccagagacta gtttcagttt ttgataagaa
      241 tgaattatta ttcattttct agtttatgaa tatatatgta actagggaag cctatttatg
      301 atttttctaa ttttttaaaa ttgttatttt agagaggtgt gaaattaaaa tgatatatat
      361 tttcgagtac tttaaaaaat aaaaacggta catcctatat taaatttttg attttagatc
      421 cttccaagtt attgacaagt attaactgca cttctcataa aaatttttca aatagatcta
      481 tttagtctgg aataggaata cttttggcaa ttttcttgag aatcatagaa gacgaatttc
      541 tgaactgctc tttgtgaagc cacgcccata tattggcatt tttgtatcca attttttgta
      601 gagtacctga atcattatca ctacatctgc taaacaatat atggaagttc tatgagaggt
      661 gaaaggtaaa tcatataaca actgtaataa catagaaatg attgtagtat atttccaatt
      721 aggctgagtt gatcttattg aagagtttga atcatagcaa ttttttcctc ttttagtatt
      781 tttcaaaaca aattttcaca atgttagatg tagtatttag cattaaggca tctgtaatct
      841 cccagaaatg ttttgggaat aaaacccttt gactctttgg tccttttttg ctccaaataa
      901 tatctgattt taacccgttc tgatctactt tttacctttc gatcgtcaca tccgcgtaca
      961 cgtttctctt gctctcaatt gcttatcttt atggcctttt cccccacttt ttgtcgacta
     1021 tcatgcggac gtggcttcct tgctccctcc cggttttttt gaaaacactt gtgaatttat
     1081 tttttgtggt caaaattacg aaacatttga attctcaatc tatttataga ccgggtgtgg
     1141 tatgttatgc catgtttttg tcattgccaa atgaacatcg atattttaat tttctgaact
     1201 taccgtaccc cctgcgttac tattttattt ttaatttgta taatttcatt gctataattt
     1261 gttgtccttg aaacattgaa gatatcgaag aaataatgac aaaaatcaga tagaacaatt
     1321 tttcctactc atcaactcac atttttctat tttctcttgt gcgaaaaaaa tagtatattg
     1381 ggaatattta tagtattgca acttttctct tagttcgcag agttgcaata ctactagatc
     1441 cctaatagag gaaatatggt gaccagcttt ttgcaattgg aaagttggga aaaaagtata
     1501 tatattaata gacttacact cttgtagttc gtagaatttt tcaggaaatc tgacacgcaa
     1561 gactttttca aaaagttttt ttttttaatt tggcaatctg ccaaaacgtt tactgaacat
     1621 tttgaaaaca tctaccagta cctttaacac aacgaaaaat tgttctaagc aatattaact
     1681 gtgataattg atgttaagac ctgaaattta atcacaaatt tcacctggaa atgacagtaa
     1741 tcggcttcta aatactgtcg acacaagtgc catacctaag attccaatcc tattaccatg
     1801 tttcccaatt tcattctctt tcgtatctct atttctcttc tttattatca attacttttt
     1861 atcagttctt cgtttgcttt aacatcaaga gcacatagcg ctctctcgtc tctctctcat
     1921 gctctttttg aacttttaca aaaaactatt tccggttgtt ttgattcttg gaatttgaaa
     1981 taattttcag tgattaaatc tagactctga ggccttgggt gctcgtcgtt ggacttttat
     2041 tgatttctag aggatccccg ggattggcca aaggacccaa aggtatgttt cgaatgatac
     2101 taacataaca tagaacattt tcaggaggac ccttggctag catgcagatc ttcgtgaaga
     2161 ccctgactgg taagaccatc actctcgaag tggagccgag tgacaccatt gagaatgtca
     2221 aggcaaagat ccaagacaag gaaggcatcc ctcctgacca gcagaggttg atctttgctg
     2281 ggaaacagct ggaagatgga cgcaccctgt ctgactacaa catccagaaa gagtccaccc
     2341 tgcacctggt cttccgtctc agaggtgtgg tggggaagct tggtcgacac ctgaaggcta
     2401 gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2461 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2521 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2581 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2641 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2701 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2761 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2821 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2881 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2941 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3001 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3061 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taggatatct
     3121 gagctccgca tcggccgctg tcatcagatc gccatctcgc gcccgtgcct ctgacttcta
     3181 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     3241 tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc
     3301 tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa
     3361 aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca caatgttctg
     3421 tgtacacttc ttatgttttt tttacttctg ataaattttt tttgaaacat catagaaaaa
     3481 accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt
     3541 cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag
     3601 tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt
     3661 tttgggggtt tcccctattg tttgtcaaga gtttcgagga cggcgttttt cttgctaaaa
     3721 tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc
     3781 agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc
     3841 cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg
     3901 tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     3961 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4021 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4081 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4141 taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4201 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5221 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5281 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5521 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5641 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5701 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5761 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5821 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5881 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5941 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6001 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6061 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6361 aggaaacagc t
//
