LOCUS       sequence-445698-        7252 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 7252)
  TITLE     Organ-specific electrophile responsivity mapping in live C. elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7252)
  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            "Unpublished"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..7252
                     /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(4826..4848)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4948..4967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5045..5063)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5131..5235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5236..5304,5305..6096)
                     /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(5454..5473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6267..6855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6756..6775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7009..7026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7143..7164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7179..7184,7185..7202,7203..7209)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7217..7233
                     /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     7222..7244
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7241..7252,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7241..7252,1..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 caaaaaaact
     4141 tcttcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt
     4201 agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc
     4261 cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt
     4321 ttttacttct gataaatttt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt
     4381 atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc
     4441 tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa
     4501 tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttgggggt ttcccctatt
     4561 gtttgtcaag agtttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca
     4621 cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag
     4681 ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat
     4741 tcccaatata ccaaacataa ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg
     4801 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4861 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4921 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4981 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt
     5041 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5101 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5221 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5761 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6061 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6121 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6181 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6241 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6361 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6421 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6481 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6541 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6601 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     6661 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6721 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6781 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6841 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6901 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7021 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7081 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7141 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7201 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ct
//
