LOCUS       sequence-238215-        6910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a C. elegan codon optimized florescent protein 
            (CemOrange2) fused to a ER localized gene (pisy-1) under the 
            intestinal specific promoter nhx-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  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: pBT3003
FEATURES             Location/Qualifiers
     source          1..6910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4484..4506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4606..4625
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4703..4721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4789..4893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4894..4962,4963..5754)
                     /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(5112..5131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5925..6513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6414..6433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6667..6684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6801..6822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6837..6842,6843..6860,6861..6867)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6875..6891
                     /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     6880..6902
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6899..6910,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6899..6910,1..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 taatttggca
     1201 atctgccaaa acgtttactg aacattttga aaacatctac cagtaccttt aacacaacga
     1261 aaaattgttc taagcaatat taactgtgat aattgatgtt aagacctgaa atttaatcac
     1321 aaatttcacc tggaaatgac agtaatcggc ttctaaatac tgtcgacaca agtgccatac
     1381 ctaagattcc aatcctatta ccatgtttcc caatttcatt ctctttcgta tctctatttc
     1441 tcttctttat tatcaattac tttttatcag ttcttcgttt gctttaacat caagagcaca
     1501 tagcgctctc tcgtctctct ctcatgctct ttttgaactt ttacaaaaaa ctatttccgg
     1561 ttgttttgat tcttggaatt tgaaataatt ttcagtgatt aaatctagag gatccccggg
     1621 attggccaaa ggacccaaag gtatgtttcg aatgatacta acataacata gaacattttc
     1681 aggaggaccc ttggctagcc tcgagaggcc tggtaccatg ggcggaaaaa tggtctccaa
     1741 gggagaggag aacaacatgg ccatcatcaa ggagttcatg cgtttcaagg tccgtatgga
     1801 gggatccgtc aacggacacg agttcgagat cgagggagag ggagagggac gtccatacga
     1861 gggattccaa accgccaagc tcaaggtcac caaggtaagt ttaaacatat atatactaac
     1921 taaccctgat tatttaaatt ttcagggagg accactccca ttcgcctggg acatcctctc
     1981 cccacacttc acctacggat ccaaggccta cgtcaagcac ccagccgaca tcccagacta
     2041 cttcaagctc tccttcccag agggattcaa gtgggagcgt gtcatgaact acgaggacgg
     2101 aggagtcgtc accgtcaccc aagactcctc cctccaagac ggagagttca tctacaaggt
     2161 aagtttaaac agttcggtac taactaacca tacatattta aattttcagg tcaagctccg
     2221 tggaaccaac ttcccatccg acggaccagt catgcaaaag aagaccatgg gatgggaggc
     2281 ctcctccgag cgtatgtacc cagaggacgg agccctcaag ggaaagatca agatgcgtct
     2341 caagctcaag gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
     2401 ggacggagga cactacacct ccgaggtcaa gaccacctac aaggccaaga agccagtcca
     2461 actcccagga gcctacatcg tcgacatcaa gctcgacatc acctcccaca acgaggacta
     2521 caccatcgtc gagcaatacg agcgtgccga gggacgtcac tccaccggag gaatggacga
     2581 actctacaag cttaacggtc ctaggagatc tgaattcgaa ccgacagccg ccgagaacaa
     2641 caacaatgtg tggctattct acccgaatct catcggttat ggacgcatag tactcgccat
     2701 tttggccatg tactacatgt cgtcagcgcc atggtgtgca atgttctggt gagttttggc
     2761 gagctggaaa tcgatttttg catcaaaaat tagctaatat ccatgtttac tactactttt
     2821 catctctttt tcactgaaat gtcgagtatt tttgcgatgc atggtaaaga aatcaaggtg
     2881 accaggtttc ttttcaattt tcccccataa ttgaagagtt acaatgccat ctgacaaggt
     2941 ttttcagaag aaaagtcctg aaaattagat tttcgtaatt aaaatttctc ataatttcag
     3001 ctatgcactc agcgccggac tcgacgcgtt tgacggatgg gcggcacgga cctacaatca
     3061 atcgagccga tttggagcaa tgctcgatca gctcaccgat cggtacgtta aaatttctca
     3121 aatatgctgg aaaattcgag aaaaaaattg gattcggtaa aattctcata ttttcagctg
     3181 cgccctgctg gccttggtga tggcgctctg caagttctac ccggatcact tgttcctgct
     3241 tcaactgtcg gccgtaatcg acatcgcatc tcactggctt caccttcacg ccactgatct
     3301 ctccggtgcc acgagccaca agcagtcttc gaattggctt ctgaacctct actacacgga
     3361 tcgattgttc ctcggattca tgtgcggcgg aaatgaggct ttctacattt tgctctacat
     3421 tcgcgcgttc tgggctggta aatatttgat ttatttttgc tataatttat taattcctct
     3481 aattttcagg accactgatc ttcaacttcc acttcatgtc gatcttccca ctcatcgcct
     3541 tcccgattgc cttcgtcaag tccgccatct ccgtcgtcca tctgtacact gccgccgaga
     3601 tggtcgttgc tcacgatgtg aagctccgtg ttgaggctgc tcagaaggcc caatgaggcg
     3661 cctaagagct ccgcatcggc cgctgtcatc agatcgccat ctcgcgcccg tgcctctgac
     3721 ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc
     3781 ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag
     3841 tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc
     3901 gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg
     3961 ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa
     4021 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat
     4081 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg
     4141 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc
     4201 tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa
     4261 tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc
     4321 agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc
     4381 cttaaatgac agaatacatt cccaatatac caaacataac tgtttctact agtcggccgt
     4441 acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4501 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4561 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4621 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4681 accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4741 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4861 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5101 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5161 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5221 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5281 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5341 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5401 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5461 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5521 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5581 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5641 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5701 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5761 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5821 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5881 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6001 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6061 ccaaatacta ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6121 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6181 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6241 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6301 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6361 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6421 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6481 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6541 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6601 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6661 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6721 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6781 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6841 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6901 ggaaacagct
//
