LOCUS       sequence-47505-e        9497 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 9497)
  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 9497)
  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: pNG2470
FEATURES             Location/Qualifiers
     source          1..9497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5407..5435)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6183..6203
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7071..7093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7193..7212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7290..7308)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7376..7480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7481..7549,7550..8341)
                     /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(7699..7718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8512..9100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9001..9020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9254..9271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9388..9409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9424..9429,9430..9447,9448..9454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9462..9478
                     /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     9467..9489
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(9486..9497,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(9486..9497,1..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 cttctccgag gccttgggtg ctcgtcgttg gacttttatt
     2041 gatttctccg tctagaggat ccccgggatt ggccaaagga cccaaaggta tgtttcgaat
     2101 gatactaaca taacatagaa cattttcagg aggacccttg gctagcatgg tcatgagcag
     2161 gacacttgta cttgtggcgc tcctcggctt cgcatatgtt tggtaagtaa cctttagaag
     2221 gcttctaagc tttcatgcct gcctaccgcc tattaaaaaa ttaaaataca aaaaacctaa
     2281 aaacaactgt gtagtttctc atttcagcga atccgctctg atcacaaatc tccctggagc
     2341 tccaatctca aacttcaagc aatattcggg ctactacaat gtgggaacca agaagaatca
     2401 tatgctccac tactggttcg tcgagtctca aagcaaccca tcaactgatc cagttcttct
     2461 ctggcttacc ggtggaccag gatgctccgg attgtcagct cttctcaccg agtggggacc
     2521 atggaatgtg agtttgaggg gagcggaaaa ttgcgcaaaa agctaacatt ttagctacaa
     2581 aatattgtgg ataaattaac atgtaaggta aaaaagaggt tccaaaaaat aggttcgtgt
     2641 tatttgaaaa atgtatagct acaatattct atagttcctc tgagtcttcg tgccctgaaa
     2701 cccaattact ccattttcct atattcttgt gaaaatatgt ttttttttat ttctaacaat
     2761 accactatca ctgataatat tccgttgttt ttacttctag ttttcatgtt tttttttctc
     2821 gcccacctac cgattttcta gggacactat atatgtcaac tttgatggct cttatctcta
     2881 ccattttcaa tttatcaaaa accgccaact acttataatg tgcatcacat tgtgatctac
     2941 actttgtagt ttacaaatgt ttgataaatt ttaaaataag cgagatacaa actattaaaa
     3001 aagtatatat tgttaccata aaaaattgtg tttaaaaaaa ttgccgttag aaattttgtt
     3061 ttaaatcgtg aatttttgaa aatgtacgac atcgaaatat tttaagaaaa gttatttctc
     3121 aagtgacatc tagaaacatg tcacttttgc tgaaatttaa gcaaacataa aaggtagaga
     3181 agcacctaca aagaagattt attaacagtt ttgtttttcc aggtcaacac cgacggagcc
     3241 acccttcgca ccaatccata ctcctggaac aagaatgcct caattctcac ccttgaagct
     3301 ccagccggag ttggatactc ttacgccaca gacaacaata ttgcaaccgg agacgaccag
     3361 acagccagtg agaattggga agccctcgtc gctttcttca atgagttccc acaatacaag
     3421 ggaaacgatt tctatgtgac tcgagagggg tacggtggaa tctatgttcc aacacttgtt
     3481 cagactattt tggatcgtca atcacaatct catatcaata ttaagggact cgcaatcgga
     3541 aacggatgtg tttcagctaa cgagggagtt gactcacttg tcaacttctt gtaccatcac
     3601 ggagttgttg atcaggttgg ggcagttttg aatatttgaa aataggaagc ctaagcctaa
     3661 ttctaagcct gaacctaagc ctcattttaa gctcaaacct aaacatacgc ctaaatctat
     3721 tttcaataaa aaaacacttc caggccaaat gggagcacat gaagacctcg tgctgccaca
     3781 acgacactga tgcctgccca tggcactcgt tcagcgagtt ctcggcctgc ggagaattcg
     3841 ttgaagctac ccaacagacc gcctggaacg gaggtctcaa cccatacaac atgtacgccg
     3901 actgcatctc caccagcgca agcttccgtt tcggaatgga gtatgagaga agattcaata
     3961 agaagtacac accagaggtt cttggaaccg ttccatgtct tgatgagagc ccagtaacca
     4021 attatttgaa ccgtcaggat gtgcgtaagg cactcggaat tccatcttct cttccagctt
     4081 ggtcgatttg caggtaagat tctaaaattt tgcattgtgt aaaaacctat atatctcaaa
     4141 atctacaagt gttaaaaaaa ggttgtcaac taacaaatta aagctcttaa agttttgcac
     4201 attttgccga caactttttt gtatgtccaa tagcaaataa gatatacctt ttccaaatat
     4261 tttactcaaa atcggtgggc gaatttgcct acatgctcca cagtaaaatg atattatttc
     4321 cagtaacgcc atctcgtacg gttacaagag acaatacgga gacatgactt cccgtgttct
     4381 gaacgccgtc aacaacaaca atctgaaaat gatgctctac aacggagacg tcgatttggc
     4441 ctgtaatgct cttatgggac agagattcac tgacaagctc ggtcttacgg taggctattt
     4501 tatgtttaaa agtttagagg cgcagatgtt ttttagtttt tcgtgatttt caatactacg
     4561 gtacctagtt ttgacacgat cgatttttta aatgaaaaag gagtgtgcgc ctttaagggg
     4621 tactgtagtt ttttctcaat ttagtctcat ttttagtaaa aagttcgaaa aacgggaatt
     4681 aaaaatttca aatatttcaa aatgcatgaa attgtgagaa aaaatcatga aacagcaaag
     4741 caaatttcaa aatttcgagc tacagtactc ctgaaaggcg caaaatcttt tacattcaaa
     4801 aaaaaatcaa tcgcttcgag accgggcacc gtatttgttt gggcataaac cacagttttt
     4861 gcttttttca ataaaaagat atctatatat tatccagacc caaagttttt ccaaacaata
     4921 cggcgccggg tctcgacacg ataaattttt gttgaatgag cgaagctgtg cgcctttaaa
     4981 gagtactgta gtttcaaact tccgttgctg cggatttttc atcgaaaatt atttttttga
     5041 aaatattact ctttaaaggc gcacatcgtt ttgcattcat aaaaattgtc gtgtcgagac
     5101 accgggtacc gtatttttaa acaataagct ctcaatcgct gcgagaccca agatttttaa
     5161 tgttttgtaa aaattccata atttaatttc cagctctcca agaagaagac ccacttcact
     5221 gtgaagggtc aaatcggtgg atacgtgaca cagtacaagg gatcccaggt gacattcgcc
     5281 actgttcgcg gagccggaca catggtacca accgacaagc cagccgtcgc cgagcacatc
     5341 atccaatcgt tcctgttcaa caaggccttc gctagcatga gtaaaggaga agaacttttc
     5401 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     5461 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
     5521 actactggaa aactacctgt tccatgggta agtttaaaca tatatatact aactaaccct
     5581 gattatttaa attttcagcc aacacttgtc actactttcg gttatggtct tcaatgcttc
     5641 gccagatacc cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt
     5701 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtaagttt
     5761 aaacagttcg gtactaacta accatacata tttaaatttt caggtgctga agtcaagttt
     5821 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     5881 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     5941 gacaaacaaa agaatggaat caaagttgta agtttaaaca tgattttact aactaactaa
     6001 tctgatttaa attttcagaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     6061 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     6121 aaccattacc tgtcctacca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     6181 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     6241 aaatgagctc cgcatcggcc gctgtcatca gatcgccatc tcgcgcccgt gcctctgact
     6301 tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc
     6361 tatttttgtt attatcaaaa aaacttcttc ttaatttctt tgttttttag cttcttttaa
     6421 gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt
     6481 cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat
     6541 gttctgtgta cacttcttat gtttttttta cttctgataa attttttttg aaacatcata
     6601 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt
     6661 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt
     6721 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt
     6781 ttgctttttg ggggtttccc ctattgtttg tcaagagttt cgaggacggc gtttttcttg
     6841 ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg
     6901 aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt
     6961 ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt
     7021 cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     7081 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     7141 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     7201 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     7261 agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa
     7321 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     7381 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7441 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7501 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7561 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7621 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7681 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7741 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7801 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7861 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7921 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7981 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8041 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8101 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8161 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8221 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8281 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8341 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8401 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8461 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     8521 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     8581 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     8641 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     8701 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     8761 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     8821 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     8881 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     8941 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     9001 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     9061 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     9121 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     9181 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     9241 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     9301 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     9361 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     9421 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     9481 tcacacagga aacagct
//
