LOCUS       sequence-238140-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a C. elegan codon optimized florescent protein 
            (CemOrange2) fused to a autophagic localized gene that is orthologus
            to mamalian LC3 (sqst-1) under the intestinal specific promoter 
            nhx-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  AUTHORS   Thomas BJ, Wight IE, Chou WYY, Moreno M, Dawson Z, Homayouni A, 
            Huang H, Kim H, Jia H, Buland JR, Wambach JA, Cole FS, Pak SC, 
            Silverman GA, Luke CJ
  TITLE     CemOrange2 fusions facilitate multifluorophore subcellular imaging 
            in C. elegans.
  JOURNAL   PLoS One. 2019 Mar 26;14(3):e0214257. doi: 
            10.1371/journal.pone.0214257. eCollection 2019.
  PUBMED    30913273
REFERENCE   2  (bases 1 to 6291)
  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: "2019-03-26"; volume: "14(3):e0214257. doi"; pages: " 
            10.1371/journal.pone.0214257. eCollection 2019"
COMMENT     Sequence Label: pBT3043
FEATURES             Location/Qualifiers
     source          1..6291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3865..3887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3987..4006
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4084..4102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4170..4274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4275..4343,4344..5135)
                     /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(4493..4512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5306..5894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5795..5814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6048..6065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6182..6203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6218..6223,6224..6241,6242..6248)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6256..6272
                     /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     6261..6283
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6280..6291,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6280..6291,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttccttcca agttattgac aagtattaac tgcacttctc
       61 ataaaaattt ttcaaataga tctatttagt ctggaatagg aatacttttg gcaattttct
      121 tgagaatcat agaagacgaa tttctgaact gctctttgtg aagccacgcc catatattgg
      181 catttttgta tccaattttt tgtagagtac ctgaatcatt gtcactacat ctgctaaaca
      241 atatatggaa gttctatgag aggtgaaagg taaatcatat aacaactgta ataacataga
      301 aatgattgta gtatatttcc aattaggctg agttgatctt attgaagagt ttgaatcata
      361 gcaatttttt cctcttttag tatttttcaa aacaaatttt cacaatgtta gatgtagtat
      421 ttagcattaa ggcatctgta atctcccaga aatgttttgg gaataaaacc ctttgactct
      481 ttggtccttt tttgctccaa ataatatctg attttaaccc gttctgatct actttttacc
      541 tttcgatcgt cacatccgcg tacacgtttc tcttgctctc aattgcttat ctttatggcc
      601 ttttccccca ctttttgtcg actatcatgc ggacgtggct tccttgctcc ctcccggttt
      661 ttttgaaaac acttgtgaat ttattttttg tggtcaaaat tacgaaacat ttgaattctc
      721 aatctattta tagaccgggt gtggtatgtt atgccatgtt tttgtcattg ccaaatgaac
      781 atcgatattt taattttctg aacttaccgt accccctgcg ttactatttt atttttaatt
      841 tgtataattt cattgctata atttgttgtc cttgaaacat tgaagatatc gaagaaataa
      901 tgacaaaaat cagatagaac aatttttcct actcatcaac tcacattttt ctattttctc
      961 ttgtgcgaaa aaaatagtat attgggaata tttatagtat tgcaactttt ctcttagttc
     1021 gcagagttgc aatactacta gatccctaat agaggaaata tggtgaccag ctttttgcaa
     1081 ttggaaagtt gggaaaaaag tatatatatt aatagactta cattcttgta gttcgtagaa
     1141 tttttcagga aatctgacac gcaagacttt ttcaaaaagt tttttttttt aatttggcaa
     1201 tctgccaaaa cgtttactga acattttgaa aacatctacc agtaccttta acacaacgaa
     1261 aaattgttct aagcaatatt aactgtgata attgatgtta agacctgaaa tttaatcaca
     1321 aatttcacct ggaaatgaca gtaatcggct tctaaatact gtcgacacaa gtgccatacc
     1381 taagattcca atcctattac catgtttccc aatttcattc tctttcgtat ctctatttct
     1441 cttctttatt atcaattact ttttatcagt tcttcgtttg ctttaacatc aagagcacat
     1501 agcgctctct cgtctctctc tcatgctctt tttgaacttt tacaaaaaac tatttccggt
     1561 tgttttgatt cttggaattt gaaataattt tcagtgatta aatctagagg atccccggga
     1621 ttggccaaag gacccaaagg tatgtttcga atgatactaa cataacatag aacattttca
     1681 ggaggaccct tggctagcct cgagaggcct ggtaccatgg gcggaaaaat ggtctccaag
     1741 ggagaggaga acaacatggc catcatcaag gagttcatgc gtttcaaggt ccgtatggag
     1801 ggatccgtca acggacacga gttcgagatc gagggagagg gagagggacg tccatacgag
     1861 ggattccaaa ccgccaagct caaggtcacc aaggtaagtt taaacatata tatactaact
     1921 aaccctgatt atttaaattt tcagggagga ccactcccat tcgcctggga catcctctcc
     1981 ccacacttca cctacggatc caaggcctac gtcaagcacc cagccgacat cccagactac
     2041 ttcaagctct ccttcccaga gggattcaag tgggagcgtg tcatgaacta cgaggacgga
     2101 ggagtcgtca ccgtcaccca agactcctcc ctccaagacg gagagttcat ctacaaggta
     2161 agtttaaaca gttcggtact aactaaccat acatatttaa attttcaggt caagctccgt
     2221 ggaaccaact tcccatccga cggaccagtc atgcaaaaga agaccatggg atgggaggcc
     2281 tcctccgagc gtatgtaccc agaggacgga gccctcaagg gaaagatcaa gatgcgtctc
     2341 aagctcaagg taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag
     2401 gacggaggac actacacctc cgaggtcaag accacctaca aggccaagaa gccagtccaa
     2461 ctcccaggag cctacatcgt cgacatcaag ctcgacatca cctcccacaa cgaggactac
     2521 accatcgtcg agcaatacga gcgtgccgag ggacgtcact ccaccggagg aatggacgaa
     2581 ctctacaagc ttaacggtcc taggagatct gaattcaagt gggcttacaa ggaggagaac
     2641 aactttgaga agcgtcgtgc cgaaggagac aagatccgca gaaagtaccc agaccgtatt
     2701 ccagtgattg ttgagaaagc accaaagtca aagctccatg acttggataa gaagaagtac
     2761 ttggtcccat ccgatcttac tgttggacag ttctacttcc tcatcagaaa acgcatccaa
     2821 cttcgtccag aagatgctct gttcttcttt gtcaacaatg tcattccaca aaccatgacc
     2881 acaatgggac aactctacca ggtaactaac cagtcgttta ttttcattta attaaccctt
     2941 ttctttatta caggaccatc acgaggaaga cttgttcctt tacatcgcct acagtgacga
     3001 aagtgtgtat ggaggagagg tcgaaaagaa ggaataaggc gcctaagagc tccgcatcgg
     3061 ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc caattactct
     3121 tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa
     3181 aaaacttctc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
     3241 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
     3301 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
     3361 gttttttact tctgataaat ttttttgaaa catcatagaa aaaaccgcac acaaaatacc
     3421 ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca cgtctgggcc
     3481 tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt ggaatttttc
     3541 aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg tttcccctat
     3601 tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca
     3661 cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag
     3721 ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat
     3781 tcccaatata ccaaacataa ctgtttctac tagtcggccg tacgggccct ttcgtctcgc
     3841 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3901 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3961 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4021 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     4081 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4141 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4201 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4261 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4321 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4381 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4441 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4501 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4561 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4621 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4681 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4741 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4801 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4861 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4921 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4981 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5041 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5101 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6001 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6061 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6121 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6181 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6241 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc t
//
