LOCUS       sequence-140488-        6419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  superfolder GFP (myo-3::sfGFP::let-858, using backbone as in L3785).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  AUTHORS   Arribere JA, Cenik ES, Jain N, Hess GT, Lee CH, Bassik MC, Fire AZ
  TITLE     Translation readthrough mitigation.
  JOURNAL   Nature. 2016 Jun 1;534(7609):719-723. doi: 10.1038/nature18308.
  PUBMED    27281202
REFERENCE   2  (bases 1 to 6419)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2016-06-1- 1"; volume: "534"; issue: "7609"; pages:
            "719-723"
COMMENT     Sequence Label: pJA327
FEATURES             Location/Qualifiers
     source          1..6419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2505..2533)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3281..3301
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3859..3881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3981..4000
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4078..4096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4164..4268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4269..4337,4338..5129)
                     /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(4487..4506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5300..5888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5789..5808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6042..6059
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6176..6197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6212..6217,6218..6235,6236..6242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6250..6266
                     /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     6255..6277
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6274..6290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6274..6290
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatccta tcgattcgcg gccgctgtac acccgggtgc cctatagtga gtcgtattgg
     2461 taccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
     2521 ttgaattaga tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg
     2581 atgcaacaaa cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
     2641 catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa
     2701 cacttgtcac tactcttaca tatggtgttc aatgcttctc gagataccca gatcatatga
     2761 aacgacatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
     2821 cattcaaaga tgacgggact tacaagacac gtaagtttaa acagttcggt actaactaac
     2881 catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata
     2941 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     3001 aatacaactt taactcacac aatgtataca tcacagcaga caaacaaaag aatggaatca
     3061 aagctgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact
     3121 tcaaaattag acacaacgtt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     3181 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     3241 ctgtcctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     3301 cagctgctgg gattacacat ggcatggacg aactatacaa aagcccacaa gctttggaat
     3361 tcgctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa
     3421 ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt
     3481 ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac attttttttt ctctttccgt
     3541 ctcccaattc gtattccgct cctctcatct gaacacaatg tgcaagttta tttatcttct
     3601 cgctttcatt tcattaggac gtggggggaa ttggtggaag ggggaaacac acaaaaggat
     3661 gatggaaatg aaataaggac acacaatatg caacaacatt caattcagaa atatggagga
     3721 aggtttaaaa gaaaacataa aaatatatag aggaggaagg aaaactagta aaaaataagc
     3781 aaagaaatta ggcgaacgat gagaattgtc ctcgcttggg ccctttcgtc tcgcgcgttt
     3841 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3901 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3961 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     4021 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc
     4081 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4141 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4201 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4261 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4321 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4381 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4441 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4501 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4561 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4621 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4681 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4741 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4801 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4861 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4921 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4981 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5041 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5101 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa
     5701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5941 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6001 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6061 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6181 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6241 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6301 ccaagctgta agtttaaaca tgatcttact aactaactat tctcatttaa attttcagag
     6361 cttaaaaatg gctgaaatca ctcacaacga tggatacgct aacaacttgg aaatgaaat
//
