LOCUS       sequence-238132-        8310 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 p62 (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 8310)
  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 8310)
  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: pBT3037
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5884..5906)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6006..6025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6103..6121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6189..6293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6294..6362,6363..7154)
                     /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(6512..6531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7325..7913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7814..7833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8067..8084
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8201..8222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8237..8242,8243..8260,8261..8267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8275..8291
                     /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     8280..8302
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8299..8310,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8299..8310,1..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 tggctagcaa aaatggctgc tgcatcatcc gctcctcacc aaatgcaagt
     1741 tcgtgtgact ttctatacaa gtcacaatcc gcaatcggtt ctcatgattt tgaagaacga
     1801 tcaggctctt caacagatct ccgagaaggc aagagaactg ttcccatcgg ctgattatcg
     1861 tctctactac ggtaagatat tatacttttg tttttgcccc gtttgctaga cgaatcaatt
     1921 gtgcaaatag agtatttgta tgccacacct atgttatgac aaaaaagtac ggtgcccggt
     1981 ctcggcacga ttgaaatttt taatggacgt tccgaggaag agtgtgcgcc tttaaacaaa
     2041 ccattttttt aaatttaaag tttaactaga tttaaaaggt cgttattttt gtaaaactca
     2101 atttaattag attatattta agatttctaa tttccctaga tatctcacag taactttttt
     2161 ccaaatcttc aaattttcag gtgatgtcaa cggaccaatc tttgagttca ccaattcgga
     2221 tcagattatg tctaagattc gaatcactac attctctcga actcctcaac tttttgttcg
     2281 cctggaaact gttccccttc gtccaacagc cctaccatca gtgagcactg aaactgccaa
     2341 aaaagcttcc gatgctgatc gtgaatcaaa gagcagatcc tcgctgaagc gtatcgagaa
     2401 caaccaggag ctcacattga agacgctgga gaagttgcaa aaggagctct caagagttag
     2461 cgctttggag acgatggttg ctgatcttca tgcaaaggaa agagaacctg ttgtcgagga
     2521 agttcagaag ccattccctc cagtgaagca ctcggcgtct tgtgatgctt gcctcggtga
     2581 catcatcgga cataggttca agtgtctcga atgtgccgat tatgatttgt gcgagaagtg
     2641 tgagagaaaa tctttacatt atgagcatgc aatgatcaga attgtggagc caaagaaaac
     2701 aatgatccca tcatatgtga catcaaactc tccgaacaat gtcttcccaa cttacatgca
     2761 gcacaagatc caacaaccac aaactatcaa ggggatcaac aagactgtta tcacgagact
     2821 cgttccttca catcaagtta atccagctcc aatgcctcaa gctccaccag tctccactcc
     2881 agttccattc cctccatctc caaccatcgg aaatggagca gttccaaaac cagcccagga
     2941 gcctcggatc ccaccatctc caacttctgc tctaccacca ccacaattct tcaatgagga
     3001 gacccgtgca gccattatta atagtgcagc gttgctcaag gaaactttca gtactttgag
     3061 cagaagcttc ttggatttgg cagatggtgc acaggccaat atggataaaa agatcattga
     3121 acgctcgacc atggaagcag gaatgaccaa attcgaagag aaaatggata atttgaaaag
     3181 acactgtgag gagaaatatc ttggaacgga aactgctgaa gctgagaaga gcgaggaatc
     3241 ttcgaagaag caagcagata gcttgtctgg accaaagatc aagaaggcta agaagaagtc
     3301 ctgctacaag aaatctgccg agagactatt cacaatgcaa cttgctgcgg cggctgctga
     3361 gcaagcccgt cttgaacaag aagccaagaa aagagctgaa gccagctctg ctgaattaca
     3421 gaggaaactt gctgagctta atgtgaagga atctcaatca aaatccgagg aaaacattgc
     3481 tgactttgtc ctatacgatc tttacggaaa ctccttggaa gctttgttga gaggtcgttc
     3541 gagctctaga atcgccgatc ctccgaccac tccaaatacc gaaccagcca ccacagagtt
     3601 agcttcaacg acctctgcaa ctccagccgt cactgctcca gcatcaattg ttccagttac
     3661 tccaactgca ccaattgttg aaactctgat cgttccacca cttccaccag tggaatcaaa
     3721 tccagaagag gttacaccaa tggtgactgc accaatctca attcactctt ctttcgagaa
     3781 catttcttcg gatttcgaga gcctcagtcc atcatgggga ggatacccag atttggaagc
     3841 tgcacaacgt ctagaagtca tcgatgataa cttactcgac ttctctgatg ttgctgttga
     3901 agcccaggaa gctcaagaag ctcaagaaga agctcgccca gagccagagc aatctctacc
     3961 accatttaat gaggaaactg aaaaggtaaa tctaaatgtc gtccatgaat cccctaatct
     4021 aacgaatttt cagacattcg atcgtctcct tgagatggga ttctcctaca acgtcactgt
     4081 tgctgcaatc agagcaaatg gatcgaatct cgagatgtgt cttcaggcgc ttcttcacct
     4141 cgagaggcct ggtaccatgg gcggaaaaat ggtctccaag ggagaggaga acaacatggc
     4201 catcatcaag gagttcatgc gtttcaaggt ccgtatggag ggatccgtca acggacacga
     4261 gttcgagatc gagggagagg gagagggacg tccatacgag ggattccaaa ccgccaagct
     4321 caaggtcacc aaggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
     4381 tcagggagga ccactcccat tcgcctggga catcctctcc ccacacttca cctacggatc
     4441 caaggcctac gtcaagcacc cagccgacat cccagactac ttcaagctct ccttcccaga
     4501 gggattcaag tgggagcgtg tcatgaacta cgaggacgga ggagtcgtca ccgtcaccca
     4561 agactcctcc ctccaagacg gagagttcat ctacaaggta agtttaaaca gttcggtact
     4621 aactaaccat acatatttaa attttcaggt caagctccgt ggaaccaact tcccatccga
     4681 cggaccagtc atgcaaaaga agaccatggg atgggaggcc tcctccgagc gtatgtaccc
     4741 agaggacgga gccctcaagg gaaagatcaa gatgcgtctc aagctcaagg taagtttaaa
     4801 catgatttta ctaactaact aatctgattt aaattttcag gacggaggac actacacctc
     4861 cgaggtcaag accacctaca aggccaagaa gccagtccaa ctcccaggag cctacatcgt
     4921 cgacatcaag ctcgacatca cctcccacaa cgaggactac accatcgtcg agcaatacga
     4981 gcgtgccgag ggacgtcact ccaccggagg aatggacgaa ctctacaagc ttaacggtcc
     5041 taggagatct gaattcggcg cctaagagct ccgcatcggc cgctgtcatc agatcgccat
     5101 ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt
     5161 tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt
     5221 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     5281 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     5341 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt
     5401 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     5461 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     5521 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     5581 attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga
     5641 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     5701 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     5761 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     5821 tgtttctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5881 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5941 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6001 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6061 cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat
     6121 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6181 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6241 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6301 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6361 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6421 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6481 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6541 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6601 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6661 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6721 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6781 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6841 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6901 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6961 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7021 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7081 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7141 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7201 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7261 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7321 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7381 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7441 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7501 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7561 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7621 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7681 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7741 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7801 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7861 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7921 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7981 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8041 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8101 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8161 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8221 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8281 gcggataaca atttcacaca ggaaacagct
//
