LOCUS       sequence-54311-e        4101 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 4101)
  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 4101)
  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: pAV1997
FEATURES             Location/Qualifiers
     source          1..4101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             138..848
                     /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(283..301)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     551..570
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1675..1697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1797..1816
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1894..1912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1980..2084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2085..2153,2154..2945)
                     /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(2303..2322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3116..3704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3605..3624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3858..3875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3992..4013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4028..4033,4034..4051,4052..4058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4066..4082
                     /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     4071..4093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4090..4101,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4090..4101,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat
       61 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      121 gaggaccctt ggctagcatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
      181 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      241 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
      301 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
      361 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
      421 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
      481 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
      541 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
      601 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
      661 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
      721 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
      781 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
      841 acaagtagga tatctgagct ccgcatcggc cgctgtcatc agatcgccat ctcgcgcccg
      901 tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg
      961 ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc
     1021 ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta
     1081 ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat
     1141 ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac
     1201 atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg
     1261 caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa
     1321 aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc
     1381 ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt
     1441 cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg
     1501 tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg
     1561 gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac
     1621 tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     1801 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     1861 aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg
     1921 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2521 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3241 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3481 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     3541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3901 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3961 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4021 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4081 aatttcacac aggaaacagc t
//
