LOCUS       sequence-208116-        9181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized Archon2 fluorescent voltage reporter in C.elegans 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9181)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 9181)
  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 Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pSF143-wArchon2-KGC-EGFP-ER2
FEATURES             Location/Qualifiers
     source          1..9181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3086..3097)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(5194..5196,5197..5907)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6750..6772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6872..6891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6973..6991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7059..7163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7164..7232,7233..8024)
                     /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(7382..7401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8195..8783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8684..8703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8937..8954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9071..9092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9107..9112,9113..9130,9131..9137)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9145..9161
                     /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     9150..9172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(9169..9181,1..4)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(9169..9181,1..4)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgaccatgat tacgccaagc ttgcatgcgc ggccgcactg actgggccgg cctctgtcac
       61 atactgctcg aatcggaggt aggtttgaaa aacatgcaat tcatttccaa aatatgacga
      121 attttgaaaa aaaaaaatag aaaagaaact caattaaatt accttatttt ttctagtacg
      181 tcaattttga cttgcactta aaatctgcat tttaaaaatg aaaattcagt gcagctgcag
      241 tgcagtgcgg ctattatttt cggcattttt gcaaaactta cacaccaccg ttataagaaa
      301 atatttgtct ttttttttaa atttttattc agtcgttatt cgaaaaagac ttacagtacc
      361 ttgctgaatt ccttccgcat tgcgctaaaa tttaccgatt gcaaaataat cttacatgct
      421 atagtaataa aaaatttaca tacttgccat ttcatttcat ttaattattt catccaattt
      481 atcttgatcg gtattactct ttgttaaaaa ttccactgtt caatttttac atttcagttg
      541 ttttaatatc gaagtgcaca agaagcgcga acataatgga caccatcgct caaaaattat
      601 gcttctagtt gtagttcctc gagcattttt gatctcaaaa gacatttcca agagcaataa
      661 ttgccatttt tgagctacaa gtaccatatc ctgctcataa ttcatgttcc accactccct
      721 aaagtttttt ctttggaatt ccccagactt tgcgcctaaa attctcgtta ttctctccaa
      781 ttcaattggc tcaattcaac tgactagatt tcagtgatgt aggtagcgcg gaactcagta
      841 ttaagagagg actcaactca gtttctaatc caatttcaac aaattctaag acaccgaaac
      901 gctgcaggtc tcgtagagcc tacgtgttga caggttcttc cttttcaatc tatttccgtc
      961 gatattccgg cgtcatttag ttgaaaatga gtgaaaaaat gcgatctgac gcgaattcac
     1021 gtgaactcga aaattcgaga atgtaatcat taagtttccg aacaccaagt tctctttcga
     1081 aaccaagtca atagaaaaat gagacaccca cgagaagaag gttggtccga aaaaagaaac
     1141 taaagaagtt atttatttct ctttttgatg tttgttgagc agaatatgat ttagttaagc
     1201 acatctgatt tcaacgccga tgaatgaggt cgtaggcgtc acgttcaatt tgatctgatt
     1261 tgttcagtgc ttacgagtag aaattctaaa tatgatgtta ctttttggtt ctgtaagatg
     1321 gagctgggta tacatgtggt gcacggcaat cgctatttgg caatttcagt catttcggca
     1381 aattcggcaa ttttggcgaa ttcccatatt ttggaaattt tggcaaatgc ggaaaatttc
     1441 aaaaacaagc aattaccaaa tttgccaaac gccgaaaaat ttaaaaaaaa accgacattt
     1501 gctggtattt tttttaatca ggatgctaat tatgtaaggt taatctataa attcttgaaa
     1561 cacacttaac tttgaaattg aaatcactga catttcgcgc attcgccgat aaaatctcta
     1621 aataaaataa tgtaaactgt gtccagtttt ggtacttgtt tagtatacct ggttcaaaat
     1681 attttccaat gcgttttcca tttatttatt ttagaaatga tcatttaatt aaaacgtaaa
     1741 attcgcattt tcaagcttag aaacattaat cctaagtcca atatggacaa aaagagtagt
     1801 gtattagtac caagtaagaa ccctaaaaaa aaaaatattc taattaaatt tttggcaaaa
     1861 tgccacattt ttcttttttt tctcaacggg gaaaagttgt gtttcttccc tttttattaa
     1921 attttttgtt ttccttaggc taagtacaac ttgaaatgga acagaaatta cataaattta
     1981 ttgctttcac attttgctct tcttcactca aatcttggac tcttgctttt cacaaaaaaa
     2041 accttataaa caaaaataag acatctgaaa aaatgatgaa cttcctgtca gtttgttcca
     2101 gtatccttct taaatttcct tgttttccac atcacaaaaa tcctacatca cgcttcgact
     2161 catttcctga cagtcatcac atcaccaaat gaatttctat ttttacaata tagttgtagt
     2221 atttttgtcg aacatatatt gttctaacta acttcttaaa aaattttgct cttgactgaa
     2281 cacaaatttc tgcgtcgttt cacgtttact tttctcaatt ttttaaaacc aaaatttaac
     2341 tgcaaacttc tgcattgttc taaaaactca caggttatct caaaaagata aatttcaaag
     2401 tgagattttt ttaaaaaaat tccggactgt tttttggttt tgatgttttt aatgaatttt
     2461 taaaattctt tattagtgcc tgaaattaac aataatttta aagtttgtca cactgcgaat
     2521 aaattaatct cacatttcgt atgtttagaa agatgcatga aactccgtcg ttcttatgaa
     2581 gttaattaag acatggctaa ttgaaagccc acatggtcat gaaaaagtaa aacttacaac
     2641 tttttactga atattaacca gtctctataa atatcccagc ttagccggaa tagggtcacg
     2701 aaaacatgaa aataaagctt gaacctattg ttatgatttt caatgccacc ttttgcagtt
     2761 taatatacac tattactact tctcagttca agttcacttc tgagacggaa cctttatttt
     2821 ctaactttct ttaatcttac tttttatgag ccgaatgttc tattttgtaa aatgatcaga
     2881 tacaagccca aagctaattt tgccaataag tgttagtaga tcaattgaat tacaatataa
     2941 tgacctgggc ggaatagatt tttgcactga tttttttcca agcttactct atgaatattc
     3001 atagatcatc ttcaaaggta gcgtgtgttg tagttgtgtt ttggcgggtt cgtgttattt
     3061 gactcagctc aagtagatgt tgtagtcttc cctcaatcaa cgcgtgggct tgaccctctt
     3121 taaattgagt ggaagccagc caatctattt gaaatctacc cccgaaatga gattcgacga
     3181 tgacacgacg tgtgtgttac gtgcgctgag aatttaggaa aaaaaagata atcttctttt
     3241 cgttagctgc gccacaacct gtgcagataa actccgcaca attgaagttc ttactggtcc
     3301 cacccgattt attcaattgt attgtgtctt tttttcttat ttctctacaa ctctaacttt
     3361 cttcaactat tctctacatc agtgcttttt tccaccatga aaaaagaaat gacgtcgttg
     3421 tttcttagga ggggcggagc ctcaactcaa tggtccgttc gaatagatcc cacccctcat
     3481 ataaatagga ctgaaaccaa agaagttttt aaattttcag cttaccgtgt ttctcaattt
     3541 tcaaaatagt cttttttgtg tctgaacggg ctgatcaatt atgttttttt ttgaatgaaa
     3601 aatcttcctc ccaagcctta aaattccttc caaagatcga aagttatgtc tattcaagtt
     3661 acttcttctt agatcaaata ttaaacaaaa aaatacgcca tcattctctc acatttagta
     3721 attcaaaatt tcgatttgtt ttcgaacttt tgaatttccc gcctaggaag tgacatgagt
     3781 gaaatctcat tagataaatg tgccactcac cgatgacact attttcaaat aagggggggg
     3841 ggtgtttcga ataccttgta cacgcgtttc cgcaaaacca cacaccccga gttcctcagc
     3901 ttcttcccct cgacccatga gccacgtcaa gatgtcagcg tgtcactttt cattgcttca
     3961 ccttctacgg taacgagaaa aggcggggaa tccaggatct ggtgtgactt cttccttctt
     4021 cgtatcttcc taaaaatgtt cgtttacttt ttagcgagtg tcatttgatg ggatgtcgtt
     4081 cttgaggaaa gaggctagaa gcttaatact ctcatatttc atattattgg catcaagacg
     4141 tttattattt aactacaata acgattgctt taacaactaa aaataataat ttcagggtta
     4201 gacaagcaac ggcgcgcctc tagaggatcc ccgggattgg ccaaaggacc caaaggtatg
     4261 tttcgaatga tactaacata acatagaaca ttttcaggag gacccttggc tagcgtcgac
     4321 ggtaccatgg tttcaattgc tcttcaagct ggatatgatc ttcttggaga tggaagacca
     4381 gaaacacttt ggcttggaat tggaacactt cttatgctta ttggaacatt ctatttcctt
     4441 gttagaggat ggggagttac agataaagat gctagagaat attatgctgt tccaattctt
     4501 gtttcaggaa ttgcttcagc tgcttatctt tcaatgttct tcggaattgg acttacagaa
     4561 gttccagttg gaggagaaat gcttgatatt tattatgcta gatatgctca ttggcttttc
     4621 tcaacaccac ttcttcttct tgatcttgct cttcttgcta aagttgatag agttattatt
     4681 ggaacacttg ttggagttga tgctcttatg attgttacag gacttattgg cgctctttca
     4741 catacagcta ttgctagata ttcatggtgg cttttctcaa caatttgtat gattgttgtt
     4801 ctttatgttc ttgctacatc acttagatca gctgctaaag aaagaggacc agaagttgct
     4861 tcaacattca atattcttac agctcttgtt cttgttcttt ggacagctta tccaattatt
     4921 tggattattg gaacagaagg agctggagtt gttggacttg gaattgaaac acttcttttc
     4981 atggttcttg atgttacatg taaagttgga ttcggattca ttcttcttag atcaagagct
     5041 attcttggag atacagaagc tccagaacca tcagctggag ctgatagatc tgctgctgat
     5101 agaccagttg ttgctgtttc aaaagctgct gctaaatcaa gaattacatc agaaggagaa
     5161 tatattccac ttgatcaaat tgatattaat gttgtttcaa aaggagaaga acttttcaca
     5221 ggagttgttc caattcttgt tgaacttgat ggagatgtta atggacataa attctcagtt
     5281 tcaggagaag gagaaggaga tgctacatat ggaaaactta cacttaaatt catttgtaca
     5341 acaggaaaac ttccagttcc atggccaaca cttgttacaa cacttacata tggagttcaa
     5401 tgtttctcaa gatatccaga tcatatgaaa caacatgatt tcttcaaatc agctatgcca
     5461 gaaggatatg ttcaagaaag aacaattttc ttcaaagatg atggaaatta taaaacaaga
     5521 gctgaagtta aattcgaagg agatacactt gttaatagaa ttgaacttaa aggaattgat
     5581 ttcaaagaag atggaaatat tcttggacat aaacttgaat ataattataa ttcacataat
     5641 gtttatatta tggctgataa acaaaaaaat ggaattaaag ttaatttcaa aattagacat
     5701 aatattgaag atggatcagt tcaacttgct gatcattatc aacaaaatac accaattgga
     5761 gatggaccag ttcttcttcc agataatcat tatctttcaa cacaatcagc tctttcaaaa
     5821 gatccaaatg aaaaaagaga tcatatggtt cttcttgaat tcgttacagc tgctggaatt
     5881 acacttggaa tggatgaact ttataaattc tgttatgaaa atgaagtttg agagctccgc
     5941 atcggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt
     6001 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     6061 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     6121 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     6181 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     6241 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     6301 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     6361 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     6421 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     6481 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     6541 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     6601 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     6661 tacattccca atataccaaa cataactgtt tctactagtc ggccgtacgg gccctttcgt
     6721 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6781 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6841 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6901 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     6961 ccttaattaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     7021 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7081 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7141 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7201 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7261 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7321 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7381 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7441 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7501 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7561 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7621 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7681 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7741 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7801 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7861 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7921 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7981 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8821 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8881 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8941 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     9001 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     9061 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     9121 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     9181 a
//
