LOCUS       sequence-238208-        6767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a C. elegan codon optimized fluorescent protein 
            (CemOrange2) fused to a lysosome localized gene (lmp-1) under the 
            intestinal specific promoter nhx-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6767)
  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 6767)
  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: pBT2999
FEATURES             Location/Qualifiers
     source          1..6767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4341..4363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4463..4482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4560..4578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4646..4750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4751..4819,4820..5611)
                     /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(4969..4988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5782..6370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6271..6290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6524..6541
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6658..6679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6694..6699,6700..6717,6718..6724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6732..6748
                     /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     6737..6759
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6756..6767,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6756..6767,1..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 aaatgttgaa atcgtttgtc atcttgtttg gtaagtgaaa
     1741 tacttcctac gcagaaatac aatagcaatc aatcaatttt cagccttcct ggcctccgca
     1801 tccgccaccg cttcgcatta ctatgtaacc aacaacaaca ccggactcac gtgcatcatc
     1861 ctcgacggag acttccagtt caatttggtc ttcaacgaga agaacaccac cgagaagttc
     1921 acggtcacat tcaacgagac agtcagtgtc gaaggagatt gcaacggagt ccgcaacaat
     1981 caatcggttc agacgttgaa tgtgagtttt tgaataacaa ctttagggca atttataact
     2041 tgaaaaaatt tccagattaa gttcaaccct gaaggacaat ctgcccgtta cgccaaagag
     2101 tgggagctcg atatcgtgtt cggatcatct agcaacgtcg cctttgagat cattgactac
     2161 accctcacca ctcagagaac ggatcttgtc ccatactttg gaaagtttgt cagagacgcc
     2221 aatgctgccg gagatgttac agcaactcag accaacgctt acaaggtaat accatttctg
     2281 ctatagcatt tggggtcttg ttaccactca ctatttcttg cctaactcaa aaattttcag
     2341 tgctccaccg ccaagcttgg actcgtcgga ggatctacca ttgacatcaa gacatccaac
     2401 gtgattgcat ttgcacagat gaacggcacc gttttcccaa ctgaccaagt ctacgaggtg
     2461 tgctatttgg atgctcgcac caacgaagtt gtcccaatcg tcgtcggagc ctgcctcgcg
     2521 gctctcgtcg tggttgtgct cgtcggatac cttatcggac gtgctcgtgc aaagagacaa
     2581 ggatatgcca gcgtcctcga gaggcctggt accatgggcg gaaaaatggt ctccaaggga
     2641 gaggagaaca acatggccat catcaaggag ttcatgcgtt tcaaggtccg tatggaggga
     2701 tccgtcaacg gacacgagtt cgagatcgag ggagagggag agggacgtcc atacgaggga
     2761 ttccaaaccg ccaagctcaa ggtcaccaag gtaagtttaa acatatatat actaactaac
     2821 cctgattatt taaattttca gggaggacca ctcccattcg cctgggacat cctctcccca
     2881 cacttcacct acggatccaa ggcctacgtc aagcacccag ccgacatccc agactacttc
     2941 aagctctcct tcccagaggg attcaagtgg gagcgtgtca tgaactacga ggacggagga
     3001 gtcgtcaccg tcacccaaga ctcctccctc caagacggag agttcatcta caaggtaagt
     3061 ttaaacagtt cggtactaac taaccataca tatttaaatt ttcaggtcaa gctccgtgga
     3121 accaacttcc catccgacgg accagtcatg caaaagaaga ccatgggatg ggaggcctcc
     3181 tccgagcgta tgtacccaga ggacggagcc ctcaagggaa agatcaagat gcgtctcaag
     3241 ctcaaggtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggac
     3301 ggaggacact acacctccga ggtcaagacc acctacaagg ccaagaagcc agtccaactc
     3361 ccaggagcct acatcgtcga catcaagctc gacatcacct cccacaacga ggactacacc
     3421 atcgtcgagc aatacgagcg tgccgaggga cgtcactcca ccggaggaat ggacgaactc
     3481 tacaagctta acggtcctag gagatctgaa ttcggcgcct aagagctccg catcggccgc
     3541 tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa
     3601 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     3661 cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg
     3721 tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc
     3781 ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt
     3841 tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat
     3901 catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc
     3961 atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc
     4021 aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt
     4081 tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     4141 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     4201 taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc
     4261 aatataccaa acataactgt ttctactagt cggccgtacg ggccctttcg tctcgcgcgt
     4321 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4381 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4441 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4501 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     4561 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4981 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5281 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6541 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6601 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6661 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6721 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
