LOCUS       sequence-238121-        7115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a C. elegan codon optimized florescent protein 
            (CemOrange2) fused to a late endosome localized gene (rab-7) under 
            the intestinal specific promoter nhx-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7115)
  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 7115)
  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: pBT3014
FEATURES             Location/Qualifiers
     source          1..7115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4689..4711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4811..4830
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4908..4926)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4994..5098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5099..5167,5168..5959)
                     /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(5317..5336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6130..6718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6619..6638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6872..6889
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7006..7027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7042..7047,7048..7065,7066..7072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7080..7096
                     /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     7085..7107
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7104..7115,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7104..7115,1..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 gaattcggcg cctcgggaac cagaaagaag
     2641 gcgctgctca aagtgatcat tctcggagat tcgggcgttg gaaaaacatc tttgatgaat
     2701 caatatgtaa atcggcgatt tagtaatcaa tataaggttt gtcaattttt ttagaaactg
     2761 aaatttttgt ttaaactcaa aattctccct gaaatataaa caaatttctc cttcaggcaa
     2821 cgattggagc cgacttcctc acacgcgacg taaacattga cgacagaacc gtcactcttc
     2881 aggtgaaaaa ttatcaaaaa tagacaaaat tttacacaaa aatttcagat ctgggataca
     2941 gccggccagg aacgtttcca atcgctcgga gtcgcttttt atcgtggagc cgattgctgt
     3001 gtgctggctt ttgacgtcac aaatgccgcc tcgttcaaat cgcttgactc gtggcgtgac
     3061 gagtttctga ttcaggctag gtacggggaa tatttagaaa aatcacattt tttctgtaaa
     3121 aaaactgaaa aattcattat ttttctctaa taaaacggaa aaatccacca tttttctcca
     3181 aaaaattcga aaaatccacg gtttttctat aaaacttcaa attcttgcag cccccgcgat
     3241 ccagaccatt tcccgtttgt cctgctcgga aacaaggtgg atttggagtc acagcgtgcc
     3301 gtttcgtcaa aacgtgctca atcttggtgc cagacgaagg gaaatattcc gtattacgag
     3361 gtttctgcca aggaagctct caacgttgaa gccgcatttt tggcaattgc tcgtgatgcg
     3421 ttggctagag aatctcaggt aaaaatgcgg cgaaattgga ttttcctgtg aaattaaaaa
     3481 atgtgaaatt tcgggcggaa attgattttt ttgtaattaa aaactagtta tatagtgaaa
     3541 atagctttaa aatcgtgaaa aaagtacgca gaaatcaaga aaaactgcca aatttcggtt
     3601 gaaaatggca taacttagcg attttgaagc taaaaatcaa agaaattggt tagaaaactg
     3661 gttttttcaa aaaatttgaa aaatctaaaa ctaaataaat atatcaaaaa tatgctgcgt
     3721 tttatgtaaa aatttgaatt tttccaacaa aaagcaaact aaaaatttta aatttttcag
     3781 gaaaccaacg atttcccaga gtttcccgac caaatccgtc tcaacccaaa tcaacagaac
     3841 cagcagaatt cgggatgcaa ttgttaataa gagctccgca tcggccgctg tcatcagatc
     3901 gccatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg
     3961 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt
     4021 tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa
     4081 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata
     4141 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat
     4201 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt
     4261 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat
     4321 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt
     4381 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc
     4441 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     4501 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     4561 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     4621 ataactgttt ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac
     4681 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4741 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4801 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     4861 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg
     4921 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4981 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5041 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5101 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5161 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5221 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5281 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5341 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5401 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5461 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5521 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5581 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5641 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5701 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5761 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5821 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5881 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5941 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6001 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6061 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6241 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6841 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6901 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6961 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7021 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7081 tgtgagcgga taacaatttc acacaggaaa cagct
//
