LOCUS       sequence-389472-        8015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of wmiRFP720-T2A-HO1 in neurons of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8015)
  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 8015)
  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-wmiRFP720-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..8015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3693..3746
                     /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(5585..5607)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5707..5726
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5808..5826)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5894..5998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5999..6067,6068..6859)
                     /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(6217..6236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7030..7618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7519..7538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7772..7789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7906..7927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7942..7947,7948..7965,7966..7972)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7980..7996
                     /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     7985..8007
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8004..8015,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8004..8015,1..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 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 aaatgtttct aataycktta aaaattggaa tattttggaa aacaatttgt
      841 tttcaattgg caatttatta attctgagct tctgagcgta aatttttttt tattttggag
      901 caattctgag atgtcttagt gcatttttcg grgcttaaat aaaggcttca ccgtctgaaa
      961 atgcaacttt tttcagatga aatattaaaa aaaaaacctc ttttattaat tttgtgctcc
     1021 aaactttccc acggtacgca aatcatgtaa tatgtacttt taaacattat ttttccaatc
     1081 aaaaatatcc aacagctttt ccttacaatt tgaaaatttt tattaaaatt tcaaaaaaat
     1141 tttttttttt tttaatttcg aattttttta aacttgttgt aacttttttc cctgtcggcg
     1201 ccaattaaca aaatacgtag aattttgcgc ttattgtttt gttagttaac aaatttttga
     1261 caactttctt atcagctgag atttaggtgt ttcagttcgc aatctgcgtc tctccttgct
     1321 ctcctctatt ttaaaacccc cataactttg ttcttgtgag agttatcaaa aaattgtcaa
     1381 ctaacaaaat atagagcaca aaaatatgca cattttgtat gttgacaatt ttttgatacc
     1441 ttttttagtt ttcgagatat cttagtttta gtttagctgt acctttttct ggcaaaaaac
     1501 ttccaaattt cggcaatttt tcggcaaatt tcccggcaaa tttcgccaag tacatcgggc
     1561 tctcagacaa gtggtctgtg agtggaagtc gataaaagta gtttacgtgc tctccgtccc
     1621 cccgcttctt ctccgtctct gcttctattt ctccattgac tagtcccact actttttacg
     1681 gggcggaggg ctctcttgca ctccagcttc ttctcttttt gtgctctttt ttgcccgtat
     1741 gtgggtgctc atttattttt ctttggtttt aaaataaatt cggtttatta ggctactcgt
     1801 aaaaatgaag aaaaaacttc aaattcaaac gacattaaaa aaaaatttaa taaaaatttt
     1861 ttgaattgct caaaagcccc acctatatat atatatagct ccgcctactt ctcgcacaga
     1921 ccccgcccac tttttgactc cgccccctta ttggcaaaag ttcttgaaaa aaataaaaat
     1981 aaaaattcaa attgaaacaa ttttttttta atgaagtagt ttttttctaa attgatgaaa
     2041 atttctaaac agtttacaag caccgcccac tttttgactc cgcccctttg ttggaaaaat
     2101 tcttgtaaaa aaaaataaaa aataaaagct tctctttcaa acgattttta gttggtcttt
     2161 gtatgaaaag tttttttcct aaaatagaaa tgtctgaata atttgcaagc tccgcccttt
     2221 tcttacccag actccgcccc tttattgact aatttttttt tgtccattta cctgaagttc
     2281 aaaaaaaaaa tttacagttt catagagtaa taatttttca aaaatctctt tttccaatgt
     2341 ttaatgtgtc attataaacc cgcccagttt taagctccgc ccatttaggc cccgcccatt
     2401 tatgacccaa atattttttt ctctattcac ccgtggattt ccctccaaca agttacccca
     2461 aaaataaaaa taaaaattcc tgcaggcgcc caccgggatc cggcgcgcct ctagaggatc
     2521 cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac
     2581 attttcagga ggacccttgg ctagcgtcga cggtaccaaa aatggccgag ggatccgtcg
     2641 cccgtcaacc agaccttctt acctgcgacg acgagccaat ccacatccca ggagccatcc
     2701 agccacacgg acttcttctt gcccttgccg ccgacatgac catcgtcgcc ggatccgaca
     2761 accttccaga gcttaccgga cttgccatcg gagcccttat cggacgttcc gccgccgacg
     2821 tcttcgactc cgagacccac aaccgcctta ccatcgccct tgccgagcca ggagccgccg
     2881 tcggagcccc aatcaccgtc ggattcacca tgcgcaaggt aagtttaaac atatatatac
     2941 taactaaccc tgattattta aattttcagg acgccggatt catcggatcc tggcaccgtc
     3001 acgaccaact tatcttcctt gagcttgagc caccacaacg cgacgtcgcc gagccacaag
     3061 ccttcttccg ccgtaccaac tccgccatcc gtcgtcttca agccgccgag acccttgagt
     3121 ccgcctgcgc cgccgccgcc caagaggtcc gcaaggtaag tttaaacagt tcggtactaa
     3181 ctaaccatac atatttaaat tttcagatca ccggattcga ccgcgtcatg atctaccgtt
     3241 tcgcctccga cttctccgga tccgtcatcg ccgaggaccg ttgcgccgag gtcgagtcca
     3301 agcttggact tcactaccca gcctccttca tcccagccca agcccgccgc ctttacacca
     3361 tcaacccagt ccgcatcatc ccagacatca actaccgccc agtcccagtc accccagacc
     3421 ttaacccagt caccggacgc ccaatcgacc tttccttcgc catccttcgt tccgtctccc
     3481 caaaccacct tgagttcatg cgcaacatcg gaatgcacgg aaccatgtcc atctccatcc
     3541 ttcgcggaga gcgtctttgg ggacttatcg tctgccacca ccgcacccca tactacgtcg
     3601 accttgacgg acgccaagcc tgcaagcgtg tcgccgagcg tcttgccacc caaatcggag
     3661 tcatggagga gggatacgac atcggatccg gagagggacg cggatccctt cttacctgcg
     3721 gagatgttga ggaaaaccca ggaccaatgg agcgcccaca accagattct atgccacaag
     3781 acctttccga ggcccttaag gaggctacca aggaggttca cacccaagct gagaacgctg
     3841 agttcatgcg caacttccaa aagggacaag ttacccgtga tggattcaag cttgtcatgg
     3901 cttccctcta ccacatctac gtcgctcttg aggaggagat cgagcgcaac aaggtaagtt
     3961 taaacatata tatactaact aaccctgatt atttaaattt tcaggagtct ccagttttcg
     4021 ctccagttta cttcccagaa gagctccacc gtaaggctgc ccttgagcaa gaccttgcct
     4081 tctggtatgg accacgctgg caagaagtta tcccatacac tccagctatg caacgctacg
     4141 ttaagcgtct tcacgaggtc ggacgtaccg agccagagct tcttgtcgct cacgcttaca
     4201 cccgttacct tggagacctt tccggaggac aagtccttaa ggtaagttta aacagttcgg
     4261 tactaactaa ccatacatat ttaaattttc agaagatcgc tcaaaaggct cttgaccttc
     4321 catcttccgg agagggactt gccttcttca ctttcccaaa catcgcttct gccactaagt
     4381 tcaagcaact ttaccgctct cgtatgaact ctcttgagat gaccccagct gtccgccaac
     4441 gtgtcatcga ggaggctaag gtaagtttaa acatgatttt actaactaac taatctgatt
     4501 taaattttca gaccgctttc cttcttaaca tccaactttt cgaagagctt caagagcttc
     4561 ttacccacga caccaaggac caatctccat cccgtgcccc aggacttcgt caacgtgcct
     4621 ccaacaaggt ccaagattcc gctccagttg agactccacg tggaaagcca ccacttaaca
     4681 cccgttccca agccccactt cttcgttggg tccttaccct ctccttcctt gtcgccaccg
     4741 tcgctgtcgg actttacgcc atgtaagagc tccgcatcgg ccgctgtcat cagatcgcca
     4801 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     4861 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     4921 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     4981 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     5041 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     5101 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     5161 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     5221 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     5281 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     5341 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     5401 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     5461 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     5521 ctgtttctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5581 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5641 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5701 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5761 acagatgcgt aaggagaaaa taccgcatca ggcggcctta attaagggcc tcgtgatacg
     5821 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6181 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6301 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6541 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6601 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6721 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6781 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6841 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6901 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6961 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7681 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7741 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7801 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     7861 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7921 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7981 tgtgagcgga taacaatttc acacaggaaa cagct
//
