LOCUS       sequence-54309-e        5223 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 5223)
  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 5223)
  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: pAV1944
FEATURES             Location/Qualifiers
     source          1..5223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1260..1970
                     /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(1405..1423)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1673..1692
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2797..2819)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2919..2938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3016..3034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3102..3206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3207..3275,3276..4067)
                     /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(3425..3444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4238..4826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4727..4746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4980..4997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5114..5135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5150..5155,5156..5173,5174..5180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5188..5204
                     /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     5193..5215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5212..5223,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5212..5223,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgct ggtggacagt aactgtctgt aataataatt
       61 actcctgacc aggttgcaat tcgagttttg ataagcataa ttataccttg tacattgtgg
      121 gttttgtgct gtggacgttt tattgtgaca tccccataag ctacaagaac aaaaatgaaa
      181 tgtaaaagta ttgaaaacgt cgtacatttt atatctgagt agtatccttt gctttaaatg
      241 tccataaaac taattttata atcaataaaa caacgtttgt aaatcaactg agtttacaag
      301 tagagacatt gagggatact ttcactatgc taaagtgata atcgaccaaa taataacttc
      361 actttggtat ttattcctgt cttataaatg ttatgtatga attaaattca tatgcatatg
      421 gctcactctg acaattttta ataatcttcc agatcaatat tgactaccga tgcgggtggt
      481 cttttgcttt gaattctgct gaactttaca ccccgaacag caatgtgtgc ttcagcctaa
      541 aaaaaagtaa gtgtgttaat cagtgccccc gattcttcat tttttgcccc tctctcccgt
      601 ttcgtcggca aaagaagaga aaataaagat aagtctcaag ataggttggt aatcgctaaa
      661 gtggttgtgt ggataagagt agcaaaatgg caggaagagc actttgcgcg cacacactgt
      721 actcattgtt ctggataaaa ttctctcgtt gtttgccgtc ggatgtctgc ctctctgcat
      781 tgagccggct tcttcactat ctttagttaa cctaaaatgc cgtttctttt ctcgtatccc
      841 cactatcccg ttgaggttct ctgctctctt cgctccctac cgccagcgag caactatccg
      901 tgggggcgcc ttgctcggaa gatggggggg aagaaagaag atttttgcta tttgcacttg
      961 agaaagagac ttttcctgcg tcgatggtta gagaacagtg tgcagacact tttcagctac
     1021 ctagaattac aattggatat ccccgcctcc caatccaccc acccagggaa aaagaagggc
     1081 tcgccgaaaa tcaaagttat ctccaggctc gcgcatccca ccgagcggtt gacttctctc
     1141 caccactttt cattttaacc ctctctagag gatccccggg attggccaaa ggacccaaag
     1201 gtatgtttcg aatgatacta acataacata gaacattttc aggaggaccc ttggctagca
     1261 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1321 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1381 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     1441 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     1501 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     1561 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1621 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1681 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1741 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1801 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1861 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1921 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtag gatatctgag
     1981 ctccgcatcg gccgctgtca tcagatcgcc atctcgcgcc cgtgcctctg acttctaagt
     2041 ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt
     2101 gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct
     2161 aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa
     2221 ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt
     2281 acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca
     2341 cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc
     2401 acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt
     2461 tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg
     2521 gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt
     2581 attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag
     2641 gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg
     2701 acagaataca ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc
     2761 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2821 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2881 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2941 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     3001 caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3061 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3121 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3181 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3241 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3301 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3361 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3421 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3481 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3541 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3601 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3661 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3721 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3781 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3841 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3901 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3961 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4021 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4081 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4141 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4381 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4621 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4801 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4861 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4921 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4981 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5041 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5101 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5161 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5221 gct
//
