LOCUS       sequence-387450-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of wmCardinal-T2A-HO1 in neurons of C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  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 7915)
  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-wmCardinal-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3593..3646
                     /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(5485..5507)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5607..5626
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5708..5726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5794..5898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5899..5967,5968..6759)
                     /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(6117..6136)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6930..7518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7419..7438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7672..7689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7806..7827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7842..7847,7848..7865,7866..7872)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7880..7896
                     /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     7885..7907
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7904..7915,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7904..7915,1..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 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 tttttaaact tgttgtaact tttttccctg tcggcgccaa
     1321 ttaacaaaat acgtagaatt ttgcgcttat tgttttgtta gttaacaaat ttttgacaac
     1381 tttcttatca gctgagattt aggtgtttca gttcgcaatc tgcgtctctc cttgctctcc
     1441 tctattttaa aacccccata actttgttct tgtgagagtt atcaaaaaat tgtcaactaa
     1501 caaaatatag agcacaaaaa tatgcacatt ttgtatgttg acaatttttt gatacctttt
     1561 ttagttttcg agatatctta gttttagttt agctgtacct ttttctggca aaaaacttcc
     1621 aaatttcggc aatttttcgg caaatttccc ggcaaatttc gccaagtaca tcgggctctc
     1681 agacaagtgg tctgtgagtg gaagtcgata aaagtagttt acgtgctctc cgtccccccg
     1741 cttcttctcc gtctctgctt ctatttctcc attgactagt cccactactt tttacggggc
     1801 ggagggctct cttgcactcc agcttcttct ctttttgtgc tcttttttgc ccgtatgtgg
     1861 gtgctcattt atttttcttt ggttttaaaa taaattcggt ttattaggct actcgtaaaa
     1921 atgaagaaaa aacttcaaat tcaaacgaca ttaaaaaaaa atttaataaa aattttttga
     1981 attgctcaaa agccccacct atatatatat atagctccgc ctacttctcg cacagacccc
     2041 gcccactttt tgactccgcc cccttattgg caaaagttct tgaaaaaaat aaaaataaaa
     2101 attcaaattg aaacaatttt tttttaatga agtagttttt ttctaaattg atgaaaattt
     2161 ctaaacagtt tacaagcacc gcccactttt tgactccgcc cctttgttgg aaaaattctt
     2221 gtaaaaaaaa ataaaaaata aaagcttctc tttcaaacga tttttagttg gtctttgtat
     2281 gaaaagtttt tttcctaaaa tagaaatgtc tgaataattt gcaagctccg cccttttctt
     2341 acccagactc cgccccttta ttgactaatt tttttttgtc catttacctg aagttcaaaa
     2401 aaaaaattta cagtttcata gagtaataat ttttcaaaaa tctctttttc caatgtttaa
     2461 tgtgtcatta taaacccgcc cagttttaag ctccgcccat ttaggccccg cccatttatg
     2521 acccaaatat ttttttctct attcacccgt ggatttccct ccaacaagtt accccaaaaa
     2581 taaaaataaa aattcctgca ggcgcccacc gggatccggc gcgcctctag aggatccccg
     2641 ggattggcca aaggacccaa aggtatgttt cgaatgatac taacataaca tagaacattt
     2701 tcaggaggac ccttggctag cgtcgacggt accaaaaatg gtctccaagg gagaggagct
     2761 tatcaaggag aacatgcaca tgaagcttta catggaggga accgtcaaca accaccactt
     2821 caagtgcacc accgagggag agggaaagcc atacgaggga acccaaaccc aacgcatcaa
     2881 ggtcgtcgag ggaggaccac ttccattcgc cttcgacatc cttgccacct gcttcatgta
     2941 cggatccaag accttcatca accacaccca ggtaagttta aacatatata tactaactaa
     3001 ccctgattat ttaaattttc agggaatccc agacttcttc aagcagtcct tcccagaggg
     3061 attcacctgg gagcgcgtca ccacctacga ggacggagga gtccttaccg tcacccaaga
     3121 cacctccctt caagacggat gccttatcta caacgtcaag cttcgtggag tcaacttccc
     3181 atccaacgga ccagtcatgc agaagaagac ccttggatgg gaggccacca ccgagaccct
     3241 ttacccagcc gacggaggac ttgagggacg ctgcgacatg gcccttaagc ttgtcggagg
     3301 aggacacctt cactgcaacc ttaaggtaag tttaaacagt tcggtactaa ctaaccatac
     3361 atatttaaat tttcagacca cctaccgctc caagaagcca gccaagaacc ttaagatgcc
     3421 aggagtctac ttcgtcgacc gccgccttga gcgtatcaag gaggccgaca acgagaccta
     3481 cgtcgagcaa cacgaggtcg ccgtcgcccg ttactgcgac cttccatcca agcttggaca
     3541 caagcttaac ggaatggacg agctttacaa gggatacgac atcggatccg gagagggacg
     3601 cggatccctt cttacctgcg gagatgttga ggaaaaccca ggaccaatgg agcgcccaca
     3661 accagattct atgccacaag acctttccga ggcccttaag gaggctacca aggaggttca
     3721 cacccaagct gagaacgctg agttcatgcg caacttccaa aagggacaag ttacccgtga
     3781 tggattcaag cttgtcatgg cttccctcta ccacatctac gtcgctcttg aggaggagat
     3841 cgagcgcaac aaggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
     3901 tcaggagtct ccagttttcg ctccagttta cttcccagaa gagctccacc gtaaggctgc
     3961 ccttgagcaa gaccttgcct tctggtatgg accacgctgg caagaagtta tcccatacac
     4021 tccagctatg caacgctacg ttaagcgtct tcacgaggtc ggacgtaccg agccagagct
     4081 tcttgtcgct cacgcttaca cccgttacct tggagacctt tccggaggac aagtccttaa
     4141 ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc agaagatcgc
     4201 tcaaaaggct cttgaccttc catcttccgg agagggactt gccttcttca ctttcccaaa
     4261 catcgcttct gccactaagt tcaagcaact ttaccgctct cgtatgaact ctcttgagat
     4321 gaccccagct gtccgccaac gtgtcatcga ggaggctaag gtaagtttaa acatgatttt
     4381 actaactaac taatctgatt taaattttca gaccgctttc cttcttaaca tccaactttt
     4441 cgaagagctt caagagcttc ttacccacga caccaaggac caatctccat cccgtgcccc
     4501 aggacttcgt caacgtgcct ccaacaaggt ccaagattcc gctccagttg agactccacg
     4561 tggaaagcca ccacttaaca cccgttccca agccccactt cttcgttggg tccttaccct
     4621 ctccttcctt gtcgccaccg tcgctgtcgg actttacgcc atgtaagagc tccgcatcgg
     4681 ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc caattactct
     4741 tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa
     4801 aaaacttctc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
     4861 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
     4921 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
     4981 gttttttact tctgataaat ttttttgaaa catcatagaa aaaaccgcac acaaaatacc
     5041 ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca cgtctgggcc
     5101 tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt ggaatttttc
     5161 aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg tttcccctat
     5221 tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca
     5281 cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag
     5341 ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat
     5401 tcccaatata ccaaacataa ctgtttctac tagtcggccg tacgggccct ttcgtctcgc
     5461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     5701 attaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5761 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5821 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5881 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5941 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6001 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6061 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6121 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6181 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6241 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6301 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6361 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6421 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6481 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6541 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6601 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6661 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6721 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6781 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6841 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6901 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6961 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7021 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7081 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7141 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7201 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7261 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7321 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7381 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7441 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7501 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7561 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7621 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7681 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7741 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7801 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7861 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagct
//
