LOCUS       sequence-387271-        8230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of wmRhubarb713-T2A-HO1 in neurons of C 
            elegan.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8230)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 8230)
  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: "Nat 
            Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pSF11-wmRhubarb713-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..8230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3908..3961
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(5800..5822)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5922..5941
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6023..6041)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6109..6213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6214..6282,6283..7074)
                     /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(6432..6451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7245..7833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7734..7753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7987..8004
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8121..8142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8157..8162,8163..8180,8181..8187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8195..8211
                     /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     8200..8222
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8219..8230,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8219..8230,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccggatcca
       61 taggtatgcc tgtcattctc tatttttaaa cgttcctgca gcagtagcac aacagagcca
      121 gcagttttcc tcttcttctc catcttcttc tgcttcttcc tctcttccca aaaaaaaaaa
      181 aaaaaaaaaa agtgaggggg ctccctgccc ccccccccct cccccccccc tcttctcctt
      241 gaagctcatc cagacgtcgc tgtcctcctc ctcctcgtcc ccggggcgct gtgtgtgtgt
      301 gtgtgtgctc cgtgagcatg tagcagcttc agcccattta aaggcgcaca ctctctgcat
      361 ctctgcgtgt ctcgacactc gttttttctc ttcttttctg ctgcacacac acacacacac
      421 acacacacgc tcatgccaga gcacttcttg ctccgggggg ctccgcgcgg agcccgagag
      481 gacctcctcc tcctttttct tcttcgcttt ttagttcttc ttgcctcggt ctcgcccctt
      541 gttttgctct ccatgtagta tatacatctg ggtgtgtgtg tgtgtgtgca cctttggagc
      601 attttttgaa cctttccaga actttggagc aacattttca gcttgttcag tgcacttttt
      661 aggctgaaaa tcctattttt aggttttcga aagttttttt ggagcatttt tcgtgcaatt
      721 taggtgcaaa attaaaaaaa agggttttgt agtgaattaa aatttttttg aaactttcgc
      781 ctttttaaaa aaatgtttct aatatcttta aaaattggaa tattttggaa aacaatttgt
      841 tttcaattgg caatttatta attctgagct tctgagcgta aatttttttt tattttggag
      901 caattctgag atgtcttagt gcatttttcg ggcatacata aaaaaaaatt tatcttaata
      961 ataaaaacct taaatttagg cttcaccgtc tggaaattca attttttcag ctgaaaaact
     1021 ttcaaaaaaa aaaaaattaa attttcacat ttattttctc tggaattttg gagcaatttc
     1081 aaagctgtct tagtgcattt ttagtacaca aatatttttt catttttctt aaattttaat
     1141 aatctaagct taaataaagg cttcaccgtc tgaaaatgca acttttttca gatgaaatat
     1201 taaaaaaaaa acctctttta ttaattttgt gctccaaact ttcccacggt acgcaaatca
     1261 tgtaatatgt acttttaaac attatttttc caatcaaaaa tatccaacag cttatactta
     1321 caatttgaaa acttttatta aaatatcaaa aaaatttttt ttgtttttaa tttcgaatta
     1381 ttttaaactg gttgtaactt ttttccctgt cggcgccaat taacaaaata cgtagaattt
     1441 tgcgcttatt gttttgttag ttaacaaatt tttgacaact ttcttatcag ctgagattta
     1501 ggtgtttcag ttcgcaatct gcgtctctcc ttgctctcct ctattttaaa acccccataa
     1561 ctttgttctt gtgagagtta tcaaaaaatt gtcaactaac aaaatataga gcacaaaaat
     1621 atgcacattt tgtatgttga caattttttg ataccttttt tagttttcga gatatcttag
     1681 ttttagttta gctgtacctt tttctggcaa aaaacttcca aatttcggca atttttcggc
     1741 aaatttcccg gcaaatttcg ccaagtacat cgggctctca gacaagtggt ctgtgagtgg
     1801 aagtcgataa aagtagttta cgtgctctcc gtccccccgc ttcttctccg tctctgcttc
     1861 tatttctcca ttgactagtc ccactacttt ttacggggcg gagggctctc ttgcactcca
     1921 gcttcttctc tttttgtgct cttttttgcc cgtatgtggg tgctcattta tttttctttg
     1981 gttttaaaat aaattcggtt tattaggcta ctcgtaaaaa tgaagaaaaa acttcaaatt
     2041 caaacgacat taaaaaaaaa tttaataaaa attttttgaa ttgctcaaaa gccccaccta
     2101 tatatatata tagctccgcc tacttctcgc acagaccccg cccacttttt gactccgccc
     2161 ccttattggc aaaagttctt gaaaaaaata aaaataaaaa ttcaaattga aacaattttt
     2221 ttttaatgaa gtagtttttt tctaaattga tgaaaatttc taaacagttt acaagcaccg
     2281 cccacttttt gactccgccc ctttgttgga aaaattcttg taaaaaaaaa taaaaaataa
     2341 aagcttctct ttcaaacgat ttttagttgg tctttgtatg aaaagttttt ttcctaaaat
     2401 agaaatgtct gaataatttg caagctccgc ccttttctta cccagactcc gcccctttat
     2461 tgactaattt ttttttgtcc atttacctga agttcaaaaa aaaaatttac agtttcatag
     2521 agtaataatt tttcaaaaat ctctttttcc aatgtttaat gtgtcattat aaacccgccc
     2581 agttttaagc tccgcccatt taggccccgc ccatttatga cccaaatatt tttttctcta
     2641 ttcacccgtg gatttccctc caacaagtta ccccaaaaat aaaaataaaa attcctgcag
     2701 gcgcccaccg ggatccggcg cgcctctaga ggatccccgg gattggccaa aggacccaaa
     2761 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggctagc
     2821 gtcgacggta ccaaaaatgg ccgagggatc cgtcgcccgt caaccagacc ttcttacctg
     2881 cgacgacgag ccaatccaca tcccaggagc catccagcca cacggacttc ttcttgccct
     2941 tgccgccgac atgaccatcg tcgccggatc cgacaacctt ccagagctta ccggacttgc
     3001 catcggagcc cttatcggac gttccgccgc cgacgtcttc gactccgaga cccacaaccg
     3061 ccttaccatc gcccttgccg agccaggagc cgccgtcgga gccccaatca ccgtcggatt
     3121 caccatgcgc aaggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
     3181 tcaggacgcc ggattcatcg gatcctggca ccgtcacgac caacttatct tccttgagct
     3241 tgagccacca caacgcgacg tcgccgagcc acaagccttc ttccgccaca ccaactccgc
     3301 catccgtcgt cttcaagccg ccgagaccct tgagtccgcc tgcgccgccg ccgcccaaga
     3361 ggtccgcaag gtaagtttaa acagttcggt actaactaac catacatatt taaattttca
     3421 gatcaccgga ttcgaccgcg tcatgatcta ccgtttcgcc tccgacttct ccggagaggt
     3481 catcgccgag gaccgttgcg ccgaggtcga gtccaacctt ggacttcact acccagcctc
     3541 caccgtccca gcccaagccc gccgccttta caccatcaac ccagtccgca tcatcccaga
     3601 catcaactac cgcccagtcc cagtcacccc agaccttaac ccagtcaccg gacgcccaat
     3661 cgacctttcc ttcgccatcc ttcgttccgt ctccccagtc caccttgagt tcatgcgcaa
     3721 catcggaatg cacggaacca tgtccatctc catccttcgc ggagagcgtc tttggggact
     3781 tatcgtctgc caccaccgca ccccatacta cgtcgacctt gacggacgtc aagcctgcga
     3841 gcttgtcgcc caagtccttg cccgtgccat cggagtcatg gaggagggat acgacatcgg
     3901 atccggagag ggacgcggat cccttcttac ctgcggagat gttgaggaaa acccaggacc
     3961 aatggagcgc ccacaaccag attctatgcc acaagacctt tccgaggccc ttaaggaggc
     4021 taccaaggag gttcacaccc aagctgagaa cgctgagttc atgcgcaact tccaaaaggg
     4081 acaagttacc cgtgatggat tcaagcttgt catggcttcc ctctaccaca tctacgtcgc
     4141 tcttgaggag gagatcgagc gcaacaaggt aagtttaaac atatatatac taactaaccc
     4201 tgattattta aattttcagg agtctccagt tttcgctcca gtttacttcc cagaagagct
     4261 ccaccgtaag gctgcccttg agcaagacct tgccttctgg tatggaccac gctggcaaga
     4321 agttatccca tacactccag ctatgcaacg ctacgttaag cgtcttcacg aggtcggacg
     4381 taccgagcca gagcttcttg tcgctcacgc ttacacccgt taccttggag acctttccgg
     4441 aggacaagtc cttaaggtaa gtttaaacag ttcggtacta actaaccata catatttaaa
     4501 ttttcagaag atcgctcaaa aggctcttga ccttccatct tccggagagg gacttgcctt
     4561 cttcactttc ccaaacatcg cttctgccac taagttcaag caactttacc gctctcgtat
     4621 gaactctctt gagatgaccc cagctgtccg ccaacgtgtc atcgaggagg ctaaggtaag
     4681 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaccg ctttccttct
     4741 taacatccaa cttttcgaag agcttcaaga gcttcttacc cacgacacca aggaccaatc
     4801 tccatcccgt gccccaggac ttcgtcaacg tgcctccaac aaggtccaag attccgctcc
     4861 agttgagact ccacgtggaa agccaccact taacacccgt tcccaagccc cacttcttcg
     4921 ttgggtcctt accctctcct tccttgtcgc caccgtcgct gtcggacttt acgccatgta
     4981 agagctccgc atcggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct
     5041 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     5101 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     5161 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     5221 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     5281 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     5341 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     5401 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     5461 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     5521 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     5581 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     5641 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     5701 aatgacagaa tacattccca atataccaaa cataactgtt tctactagtc ggccgtacgg
     5761 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5821 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5881 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5941 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6001 catcaggcgg ccttaattaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6061 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6421 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6721 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7981 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     8041 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     8101 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8161 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8221 ggaaacagct
//
