LOCUS       sequence-387327-        7947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of wmiRFP680-T2A-HO1 in neurons of C elegan.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7947)
  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 7947)
  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-wmiRFP680-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..7947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3625..3678
                     /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(5517..5539)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5639..5658
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5740..5758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5826..5930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5931..5999,6000..6791)
                     /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(6149..6168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6962..7550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7451..7470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7704..7721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7838..7859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7874..7879,7880..7897,7898..7904)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7912..7928
                     /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     7917..7939
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7936..7947,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7936..7947,1..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 artgaggggg cycmcysccc 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 aatttgtttt caattggcaa tttattaatt ctgagcttct gagcgtaaat
      841 ttttttttat tttggagcaa ttctgagatg tcttagtgat ttaggcttca ccgtctggaa
      901 attcaatttt ttcagctgaa aaactttcaa aaaaaaaaaa attaaatttt cacatttatt
      961 ttctctggaa ttttggagca atttcaaagc tgtctccgtc tgaaaatgca acttttttca
     1021 gatgaaatat taaaaaaaaa acctctttta ttaattttgt gctccaaact ttcccacggt
     1081 acgcaaatca tgtaatatgt actttaactt gttgtaactt ttttccctgt cggcgcsaat
     1141 taacaaaata cgtagaattt tgcgcttatt gttttgttag ttaacaaatt tttgacaact
     1201 ttcttatcag ctgagattta ggtgtttcag ttcgcaatct gcgtctctcc ttgctctcct
     1261 ctattttaaa acccccataa ctttgttctt gtgagagtta tcaaaaaatt gtcaactaac
     1321 aaaatataga gcacaaaaat atgcacattt tgtatgttga caattttttg ataccttttt
     1381 tagttttcga gatatcttag ttttagttta gctgtacctt tttctggcaa aaaacttcca
     1441 aatttcggca atttttcggc aaatttcccg gcaaatttcg ccaagtacat cgggctctca
     1501 gacaagtggt ctgtgagtgg aagtcgataa aagtagttta cgtgctctcc gtccccccgc
     1561 ttcttctccg tctctgcttc tatttctcca ttgactagtc ccactacttt ttacggggcg
     1621 gagggctctc ttgcactcca gcttcttctc tttttgtgct cttttttgcc cgtatgtggg
     1681 tgctcattta tttttctttg gttttaaaat aaattcggtt tattaggcta ctcgtaaaaa
     1741 tgaagaaaaa acttcaaatt caaacgacat taaaaaaaaa tttaataaaa attttttgaa
     1801 ttgctcaaaa gccccaccta tatatatata tagctccgcc tacttctcgc acagaccccg
     1861 cccacttttt gactccgccc ccttattggc aaaagttctt gaaaaaaata aaaataaaaa
     1921 ttcaaattga aacaattttt ttttaatgaa gtagtttttt tctaaattga tgaaaatttc
     1981 taaacagttt acaagcaccg cccacttttt gacgcccctt tgttggaaaa attcttgtaa
     2041 aaaaaaataa aaaataaaag cttctctttc aaacgatttt tagttggtct ttgtatgaaa
     2101 agtttttttc ctaaaataga aatgtctgaa taatttgcaa gctccgccct tttcttaccc
     2161 agactccgcc cctttattga ctaatttttt tttgtccatt tacctgaagt tcaaaaaaaa
     2221 aatttacagt ttcatagagt aataattttt caaaaatctc tttttccaat gtttaatgtg
     2281 tcattataaa cccgcccagt tttaagctcc gcccatttag gccccgccca tttatgaccc
     2341 aaatattttt ttctctattc acccgtggat ttccctccaa caagttaccc caaaaataaa
     2401 aataaaaatt cctgcaggcg cccaccggga tccggcgcgc ctctagagga tccccgggat
     2461 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
     2521 gaggaccctt ggctagcgtc gacggtacca aaaatggccg agggatccgt cgcccgtcaa
     2581 ccagaccttc ttacctgcga cgacgagcca atccacatcc caggagccat ccagccacac
     2641 ggacttcttc ttgcccttgc cgccgacatg accatcgtcg ccggatccga caaccttcca
     2701 gagcttaccg gacttgccat cggagccctt atcggacgtt ccgccgccga cgtcttcgac
     2761 tccgagaccc acaaccgcct taccatcgcc cttgccgagc caggagccgc cgtcggagcc
     2821 ccaatcaccg tcggattcac catgcgcaag gtaagtttaa acatatatat actaactaac
     2881 cctgattatt taaattttca ggacgccgga ttcatcggat cctggcaccg tcacgaccaa
     2941 cttatcttcc ttgagcttga gccaccacaa cgcgacgtcg ccgagccaca agccttcttc
     3001 cgccgtacca actccgccat ccgtcgtctt caagccgccg agacccttga gtccgcctgc
     3061 gccgccgccg cccaagaggt ccgcaaggta agtttaaaca gttcggtact aactaaccat
     3121 acatatttaa attttcagat caccggattc gaccgcgtca tgatctaccg tttcgcctcc
     3181 gacttctccg gagaggtcat cgccgaggac cgttgcgccg aggtcgagtc caagcttgga
     3241 cttcactacc cagcctccac cgtcccagcc caagcccgcc gcctttacac catcaaccca
     3301 gtccgcatca tcccagacat caactaccgc ccagtcccag tcaccccaga ccttaaccca
     3361 gtcaccggac gcccaatcga cctttccttc gccatccttc gttccgtctc cccatgccac
     3421 cttgagttca tgcgcaacat cggaatgcac ggaaccatgt ccatctccat ccttcgcgga
     3481 gagcgtcttt ggggacttat cgtctgccac caccgcaccc catactacgt cgaccttgac
     3541 ggacgccaag cctgcaagcg tgtcgccgag cgtcttgcca cccaaatcgg agtcatggag
     3601 gagggatacg acatcggatc cggagaggga cgcggatccc ttcttacctg cggagatgtt
     3661 gaggaaaacc caggaccaat ggagcgccca caaccagatt ctatgccaca agacctttcc
     3721 gaggccctta aggaggctac caaggaggtt cacacccaag ctgagaacgc tgagttcatg
     3781 cgcaacttcc aaaagggaca agttacccgt gatggattca agcttgtcat ggcttccctc
     3841 taccacatct acgtcgctct tgaggaggag atcgagcgca acaaggtaag tttaaacata
     3901 tatatactaa ctaaccctga ttatttaaat tttcaggagt ctccagtttt cgctccagtt
     3961 tacttcccag aagagctcca ccgtaaggct gcccttgagc aagaccttgc cttctggtat
     4021 ggaccacgct ggcaagaagt tatcccatac actccagcta tgcaacgcta cgttaagcgt
     4081 cttcacgagg tcggacgtac cgagccagag cttcttgtcg ctcacgctta cacccgttac
     4141 cttggagacc tttccggagg acaagtcctt aaggtaagtt taaacagttc ggtactaact
     4201 aaccatacat atttaaattt tcagaagatc gctcaaaagg ctcttgacct tccatcttcc
     4261 ggagagggac ttgccttctt cactttccca aacatcgctt ctgccactaa gttcaagcaa
     4321 ctttaccgct ctcgtatgaa ctctcttgag atgaccccag ctgtccgcca acgtgtcatc
     4381 gaggaggcta aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     4441 cagaccgctt tccttcttaa catccaactt ttcgaagagc ttcaagagct tcttacccac
     4501 gacaccaagg accaatctcc atcccgtgcc ccaggacttc gtcaacgtgc ctccaacaag
     4561 gtccaagatt ccgctccagt tgagactcca cgtggaaagc caccacttaa cacccgttcc
     4621 caagccccac ttcttcgttg ggtccttacc ctctccttcc ttgtcgccac cgtcgctgtc
     4681 ggactttacg ccatgtaaga gctccgcatc ggccgctgtc atcagatcgc catctcgcgc
     4741 ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct
     4801 gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt
     4861 agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc
     4921 gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa
     4981 aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga
     5041 aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga
     5101 ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt
     5161 gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt
     5221 ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt
     5281 tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt
     5341 gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct
     5401 atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat aactgtttct
     5461 actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     5521 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5581 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5641 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     5701 gtaaggagaa aataccgcat caggcggcct taattaaggg cctcgtgata cgcctatttt
     5761 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5821 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5881 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5941 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6001 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6061 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6121 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6181 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6241 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6301 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6361 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6421 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6481 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6541 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6601 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6661 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6721 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6781 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6841 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6901 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6961 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7021 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7081 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7141 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7201 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7261 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7321 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7381 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7441 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7501 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7561 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7621 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7681 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7741 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7801 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7861 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7921 gataacaatt tcacacagga aacagct
//
