LOCUS       sequence-459063-        7244 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For organ-specific expression of separated mCherry and UltraID in 
            the body-wall muscle and neighboring vulval muscle cells of C. 
            elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7244)
  TITLE     Organ-specific electrophile responsivity mapping in live C. elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7244)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pWormgate2-myo-3p-mCherry-P2A-flag-UltraID
FEATURES             Location/Qualifiers
     source          1..7244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3397..3453
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3460..3483
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4818..4840)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4940..4959
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5037..5055)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5123..5227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5228..5296,5297..6088)
                     /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(5446..5465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6259..6847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6748..6767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7001..7018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7135..7156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7171..7176,7177..7194,7195..7201)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7209..7225
                     /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     7214..7236
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7233..7244,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7233..7244,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttagtgatt atagtctctg ttttcgttaa ttttgaattt
       61 tgcttgataa ggctgcaaca aagatcaggt tgacatatat tttcagtaat ttattttaac
      121 ctgtactcta tcactgccgg ctataataag ttcttgaata aaataatttt cccgacaaaa
      181 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      241 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      301 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      361 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      421 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      481 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      541 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      601 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      661 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      721 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      781 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      841 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      901 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      961 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
     1021 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
     1081 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1141 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1201 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1261 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1321 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1381 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1441 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1501 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1561 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1621 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1681 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1741 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1801 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1861 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1921 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1981 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     2041 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     2101 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2161 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2221 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2281 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2341 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2401 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2461 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2521 gaagctagca tggtctcaaa gggtgaagaa gataacatgg caattattaa agagtttatg
     2581 cgtttcaagg tgcatatgga gggatctgtc aatgggcatg agtttgaaat tgaaggtgaa
     2641 ggagaaggcc gaccatatga gggaacacaa accgcaaaac taaaggtaag tttaaacata
     2701 tatatactaa ctaaccctga ttatttaaat tttcaggtaa ctaaaggcgg accattacca
     2761 ttcgcctggg acatcctctc tccacagttc atgtatggaa gtaaagctta tgttaaacat
     2821 ccggcagata taccagatta tttgaaactt tcattcccgg agggttttaa gtgggaacgc
     2881 gtaatgaatt ttgaagacgg aggagttgtt acagtgacgc aagactcaag gtaagtttaa
     2941 acagttcggt actaactaac catacatatt taaattttca gcctccaaga tggagaattt
     3001 atttataaag tcaaacttcg aggaacgaat ttcccctcgg atggacctgt tatgcagaag
     3061 aagactatgg gatgggaagc ttcaagtgaa agaatgtacc ctgaagacgg tgctcttaag
     3121 ggagagatta aacaacgtct taaattgaaa gatggaggac attacgatgc tgaggtaagt
     3181 ttaaacatga ttttactaac taactaatct gatttaaatt ttcaggtgaa gacaacttac
     3241 aaagccaaaa aaccagttca gctgccagga gcgtacaatg ttaatattaa actggatatc
     3301 acctcccaca acgaggatta cactatcgtt gagcaatatg aaagagctga agggcggcac
     3361 tcgacaggtg gcatggatga attgtataag agcggagcta ctaacttcag cctgctgaag
     3421 caggctggag acgtggagga gaaccctgga cctggcagcg attataaaga cgatgacgac
     3481 aaggacttca agaacctgat ctggctgaag gaggtggaca gcacccagga gagactgaag
     3541 gagtggaacg tgtcctacgg caccgccctg gtggccgaca gacagaccaa gggcagaggc
     3601 ggcccgggca gaaagtggct gagccaggag ggcggcctgt acttcagctt cctgctgaac
     3661 cccaaggagt tcgagaacct gctgcagctg cccctggtgc tgggcctgag cgtgagcgag
     3721 gccctggagg agatcaccga gatccccttc agcctgaagt ggcccaacga cgtgtacttc
     3781 caggagaaga aggtgagcgg cgtgctgtgc gagctgagca aggacaagct gatcgtgggc
     3841 atcggcatca acgtgaacca gagagagatc cccgaggaga tcaaggacag agccaccacc
     3901 ctgtacgaga tcaccggcaa ggactgggac agaaaggagg tgctgctgaa ggtgctgaag
     3961 agaatcagcg agaacctgaa gaagttcaag gagaagtaag agctccgcat cggccgctgt
     4021 catcagatcg ccatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat
     4081 ccctacatgc tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt
     4141 ctcttaattt ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag
     4201 attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca
     4261 ttaataataa ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt
     4321 acttctgata aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat
     4381 atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg
     4441 acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag
     4501 tgaaagttta tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt
     4561 caagatttcg aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca
     4621 agaaagatcg gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa
     4681 tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat
     4741 ataccaaaca taactgtttc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc
     4801 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4861 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4921 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4981 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct
     5041 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5101 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5161 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5221 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5281 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5341 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5401 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5461 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5521 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5581 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5641 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5701 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5761 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5821 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5881 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5941 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6001 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6061 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6121 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6241 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6301 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6361 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6421 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6481 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6541 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6601 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6661 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6721 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6781 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6841 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6901 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6961 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7021 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     7081 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     7141 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     7201 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agct
//
