LOCUS       sequence-387500-        8217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of wiRFP713-T2A-HO1 in neurons of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8217)
  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 8217)
  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-wiRFP713-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3895..3948
                     /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(5787..5809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5909..5928
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6010..6028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6096..6200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6201..6269,6270..7061)
                     /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(6419..6438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7232..7820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7721..7740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7974..7991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8108..8129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8144..8149,8150..8167,8168..8174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8182..8198
                     /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     8187..8209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8206..8217,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8206..8217,1..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 ctcactgccc ccccccctct tctccttgaa gctcatccag
      241 acgtcgctgt cctcctcctc ctcgtccccg gggcgctgtg tgtgtgtgtg tgtgctccgt
      301 gagcatgtag cagcttcagc ccatttaaag gcgcacactc tctgcatctc tgcgtgtctc
      361 gacactcgtt ttttctcttc ttttctgctg cacacacaca cacacacaca cacacgctca
      421 tgccagagca cttcttgctc cggggggctc cgcgcggagc ccgagaggac ctcctcctcc
      481 tttttcttct tcgcttttta gttcttcttg cctcggtctc gccccttgtt ttgctctcca
      541 tgtagtatat acatctgggt gtgtgtgtgt gtgtgcacct ttggagcatt ttttgaacct
      601 ttccagaact ttggagcaac attttcagct tgttcagtgc actttttagg ctgaaaatcc
      661 tatttttagg ttttcgaaag tttttttgga gcatttttcg tgcaatttag gtgcaaaatt
      721 aaaaaaaagg gttttgtagt gaattaaaat ttttttgaaa ctttcgcctt tttaaaaaaa
      781 tgtttctaat atctttaaaa attggaatat tttggaaaac aatttgtttt caattggcaa
      841 tttattaatt ctgagcttct gagcgtaaat ttttttttat tttggagcaa ttctgagatg
      901 tcttagtgca tttttcgggc atacataaaa aaaaatttat cttaataata aaaaccttaa
      961 atttaggctt caccgtctgg aaattcaatt ttttcagctg aaaaactttc aaaaaaaaaa
     1021 aaattaaatt ttcacattta ttttctctgg aattttggag caatttcaaa gctgtcttag
     1081 tgcattttta gtacacaaat attttttcat ttttcttaaa ttttaataat ctaagcttaa
     1141 ataaaggctt caccgtctga aaatgcaact tttttcagat gaaatattaa aaaaaaaacc
     1201 tcttttatta attttgtgct ccaaactttc ccacggtacg caaatcatgt aatatgtact
     1261 tttaaacatt atttttccaa tcaaaaatat ccaacagctt ttccttacaa tttgaaaatt
     1321 tttattaaaa tttcaaaaaa attttttttt tttttaattt cgaatttttt taaacttgtt
     1381 gtaacttttt tccctgtcgg cgccaattaa caaaatacgt agaattttgc gcttattgtt
     1441 ttgttagtta acaaattttt gacaactttc ttatcagctg agatttaggt gtttcagttc
     1501 gcaatctgcg tctctccttg ctctcctcta ttttaaaacc cccataactt tgttcttgtg
     1561 agagttatca aaaaattgtc aactaacaaa atatagagca caaaaatatg cacattttgt
     1621 atgttgacaa ttttttgata ccttttttag ttttcgagat atcttagttt tagtttagct
     1681 gtaccttttt ctggcaaaaa acttccaaat ttcggcaatt tttcggcaaa tttcccggca
     1741 aatttcgcca agtacatcgg gctctcagac aagtggtctg tgagtggaag tcgataaaag
     1801 tagtttacgt gctctccgtc cccccgcttc ttctccgtct ctgcttctat ttctccattg
     1861 actagtccca ctacttttta cggggcggag ggctctcttg cactccagct tcttctcttt
     1921 ttgtgctctt ttttgcccgt atgtgggtgc tcatttattt ttctttggtt ttaaaataaa
     1981 ttcggtttat taggctactc gtaaaaatga agaaaaaact tcaaattcaa acgacattaa
     2041 aaaaaaattt aataaaaatt ttttgaattg ctcaaaagcc ccacctatat atatatatag
     2101 ctccgcctac ttctcgcaca gaccccgccc actttttgac tccgccccct tattggcaaa
     2161 agttcttgaa aaaaataaaa ataaaaattc aaattgaaac aatttttttt taatgaagta
     2221 gtttttttct aaattgatga aaatttctaa acagtttaca agcaccgccc actttttgac
     2281 tccgcccctt tgttggaaaa attcttgtaa aaaaaaataa aaaataaaag cttctctttc
     2341 aaacgatttt tagttggtct ttgtatgaaa agtttttttc ctaaaataga aatgtctgaa
     2401 taatttgcaa gctccgccct tttcttaccc agactccgcc cctttattga ctaatttttt
     2461 tttgtccatt tacctgaagt tcaaaaaaaa aatttacagt ttcatagagt aataattttt
     2521 caaaaatctc tttttccaat gtttaatgtg tcattataaa cccgcccagt tttaagctcc
     2581 gcccatttag gccccgccca tttatgaccc aaatattttt ttctctattc acccgtggat
     2641 ttccctccaa caagttaccc caaaaataaa aataaaaatt cctgcaggcg cccaccggga
     2701 tccggcgcgc ctctagagga tccccgggat tggccaaagg acccaaaggt atgtttcgaa
     2761 tgatactaac ataacataga acattttcag gaggaccctt ggctagcgtc gacggtacca
     2821 aaaatggccg agggatccgt cgcccgtcaa ccagaccttc ttacctgcga cgacgagcca
     2881 atccacatcc caggagccat ccagccacac ggacttcttc ttgcccttgc cgccgacatg
     2941 accatcgtcg ccggatccga caaccttcca gagcttaccg gacttgccat cggagccctt
     3001 atcggacgtt ccgccgccga cgtcttcgac tccgagaccc acaaccgcct taccatcgcc
     3061 cttgccgagc caggagccgc cgtcggagcc ccaatcaccg tcggattcac catgcgcaag
     3121 gtaagtttaa acatatatat actaactaac cctgattatt taaattttca ggacgccgga
     3181 ttcatcggat cctggcaccg tcacgaccaa cttatcttcc ttgagcttga gccaccacaa
     3241 cgcgacgtcg ccgagccaca agccttcttc cgccgtacca actccgccat ccgtcgtctt
     3301 caagccgccg agacccttga gtccgcctgc gccgccgccg cccaagaggt ccgcaaggta
     3361 agtttaaaca gttcggtact aactaaccat acatatttaa attttcagat caccggattc
     3421 gaccgcgtca tgatctaccg tttcgcctcc gacttctccg gagaggtcat cgccgaggac
     3481 cgttgcgccg aggtcgagtc caagcttgga cttcactacc cagcctccac cgtcccagcc
     3541 caagcccgcc gcctttacac catcaaccca gtccgcatca tcccagacat caactaccgc
     3601 ccagtcccag tcaccccaga ccttaaccca gtcaccggac gcccaatcga cctttccttc
     3661 gccatccttc gttccgtctc cccagtccac cttgagttca tgcgcaacat cggaatgcac
     3721 ggaaccatgt ccatctccat ccttcgcgga gagcgtcttt ggggacttat cgtctgccac
     3781 caccgcaccc catactacgt cgaccttgac ggacgtcaag cctgcgagct tgtcgcccaa
     3841 gtccttgcct ggcaaatcgg agtcatggag gagggatacg acatcggatc cggagaggga
     3901 cgcggatccc ttcttacctg cggagatgtt gaggaaaacc caggaccaat ggagcgccca
     3961 caaccagatt ctatgccaca agacctttcc gaggccctta aggaggctac caaggaggtt
     4021 cacacccaag ctgagaacgc tgagttcatg cgcaacttcc aaaagggaca agttacccgt
     4081 gatggattca agcttgtcat ggcttccctc taccacatct acgtcgctct tgaggaggag
     4141 atcgagcgca acaaggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat
     4201 tttcaggagt ctccagtttt cgctccagtt tacttcccag aagagctcca ccgtaaggct
     4261 gcccttgagc aagaccttgc cttctggtat ggaccacgct ggcaagaagt tatcccatac
     4321 actccagcta tgcaacgcta cgttaagcgt cttcacgagg tcggacgtac cgagccagag
     4381 cttcttgtcg ctcacgctta cacccgttac cttggagacc tttccggagg acaagtcctt
     4441 aaggtaagtt taaacagttc ggtactaact aaccatacat atttaaattt tcagaagatc
     4501 gctcaaaagg ctcttgacct tccatcttcc ggagagggac ttgccttctt cactttccca
     4561 aacatcgctt ctgccactaa gttcaagcaa ctttaccgct ctcgtatgaa ctctcttgag
     4621 atgaccccag ctgtccgcca acgtgtcatc gaggaggcta aggtaagttt aaacatgatt
     4681 ttactaacta actaatctga tttaaatttt cagaccgctt tccttcttaa catccaactt
     4741 ttcgaagagc ttcaagagct tcttacccac gacaccaagg accaatctcc atcccgtgcc
     4801 ccaggacttc gtcaacgtgc ctccaacaag gtccaagatt ccgctccagt tgagactcca
     4861 cgtggaaagc caccacttaa cacccgttcc caagccccac ttcttcgttg ggtccttacc
     4921 ctctccttcc ttgtcgccac cgtcgctgtc ggactttacg ccatgtaaga gctccgcatc
     4981 ggccgctgtc atcagatcgc catctcgcgc ccgtgcctct gacttctaag tccaattact
     5041 cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc
     5101 aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa
     5161 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc
     5221 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt
     5281 atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata
     5341 ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg
     5401 cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt
     5461 tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg ggtttcccct
     5521 attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag
     5581 cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga
     5641 agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac
     5701 attcccaata taccaaacat aactgtttct actagtcggc cgtacgggcc ctttcgtctc
     5761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5881 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5941 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcggcct
     6001 taattaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6061 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
