LOCUS       sequence-238211-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a C. elegan codon optimized florescent protein 
            (CemOrange2) fused to a N-terminal (sv-40) and C-terminal (egl-13) 
            nuclear localization signal under the intestinal specific promoter 
            nhx-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  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 5969)
  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: pBT3047
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1735..1755
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3543..3565)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3665..3684
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3762..3780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3848..3952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3953..4021,4022..4813)
                     /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(4171..4190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4984..5572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5473..5492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5726..5743
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5860..5881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5896..5901,5902..5919,5920..5926)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5934..5950
                     /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     5939..5961
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5958..5969,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5958..5969,1..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 ggccccaaag
     1741 aagaagcgta aggtcgtctc caagggagag gagaacaaca tggccatcat caaggagttc
     1801 atgcgtttca aggtccgtat ggagggatcc gtcaacggac acgagttcga gatcgaggga
     1861 gagggagagg gacgtccata cgagggattc caaaccgcca agctcaaggt caccaaggta
     1921 agtttaaaca tatatatact aactaaccct gattatttaa attttcaggg aggaccactc
     1981 ccattcgcct gggacatcct ctccccacac ttcacctacg gatccaaggc ctacgtcaag
     2041 cacccagccg acatcccaga ctacttcaag ctctccttcc cagagggatt caagtgggag
     2101 cgtgtcatga actacgagga cggaggagtc gtcaccgtca cccaagactc ctccctccaa
     2161 gacggagagt tcatctacaa ggtaagttta aacagttcgg tactaactaa ccatacatat
     2221 ttaaattttc aggtcaagct ccgtggaacc aacttcccat ccgacggacc agtcatgcaa
     2281 aagaagacca tgggatggga ggcctcctcc gagcgtatgt acccagagga cggagccctc
     2341 aagggaaaga tcaagatgcg tctcaagctc aaggtaagtt taaacatgat tttactaact
     2401 aactaatctg atttaaattt tcaggacgga ggacactaca cctccgaggt caagaccacc
     2461 tacaaggcca agaagccagt ccaactccca ggagcctaca tcgtcgacat caagctcgac
     2521 atcacctccc acaacgagga ctacaccatc gtcgagcaat acgagcgtgc cgagggacgt
     2581 cactccaccg gaggaatgga cgaactctac aagcttaacg gtatgagccg tagacgaaaa
     2641 gcgaatccga caaaactgag tgaaaacgcg aagaagcttg ccaaggaagt tgaaaatcct
     2701 aggagatctg aattcggcgc ctaagagctc cgcatcggcc gctgtcatca gatcgccatc
     2761 tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta catgctcttt
     2821 ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt aatttctttg
     2881 ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca aaaatagaat
     2941 taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat aataattcta
     3001 tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc tgataaattt
     3061 ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt
     3121 ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct
     3181 catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa
     3241 ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga tttcgaggac
     3301 ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa gatcggaaga
     3361 aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga aagtggagta
     3421 gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc aaacataact
     3481 gtttctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3541 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3601 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3661 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     3721 agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt
     3781 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5281 taaggcgcag cggtcgggat gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5941 cggataacaa tttcacacag gaaacagct
//
