LOCUS       sequence-350005-        9626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prgef-1 GCaMP6::3xNLS::mNeptune.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9626)
  AUTHORS   Susoy V, Hung W, Witvliet D, Whitener JE, Wu M, Park CF, Graham BJ, 
            Zhen M, Venkatachalam V, Samuel ADT
  TITLE     Natural sensory context drives diverse brain-wide activity during C.
            elegans mating.
  JOURNAL   Cell. 2021 Sep 30;184(20):5122-5137.e17. doi: 
            10.1016/j.cell.2021.08.024. Epub 2021 Sep 16.
  PUBMED    34534446
REFERENCE   2  (bases 1 to 9626)
  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"; doi: 
            "10.1016/j.cell.2021.08.024"; journalName: "Cell"; date: 
            "2021-09-30- 30"; volume: "184"; issue: "20"; pages: "5122-5137.e17"
COMMENT     Sequence Label: pJH3971
FEATURES             Location/Qualifiers
     source          1..9626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3945..3962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(4002..4010,4011..4025)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             4053..4109
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             5478..5498
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5523..5543
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5565..5585
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7394..7416)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7516..7535
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7613..7631)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7699..7803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7804..7872,7873..8664)
                     /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(8022..8041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8835..9423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9324..9343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9577..9594
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
      241 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
      301 ggatacgcta acaacttgga aatgaaataa gcttgcatgc ctgcaggtcg acgcttgcat
      361 gcctgcagcg tttccgatac ccccttatat cagcacacat tcagtcatca taccacaaaa
      421 tgttgtgata gtgataggca aaactaccgt atttcctcta ttaatcttgc atgcaagact
      481 aattttcgat taacccgtag gggtgcaaga ctaatagaga ctgcaagact attagaggct
      541 gaaatactaa ttttcgtatg ctcaataatt ttggaaattg gcctattttt tgtagaaact
      601 tgataccgtt taaacaagga aaaatacaca ctttttaata tttcatcaat aatttgaacg
      661 attttgtgat tttaagttca atttgcccaa aaaaagacaa tttttctgac gttaccgcaa
      721 agtaatgctg gtcaggcaaa aagagcggtg caaaaatata agagactgca atactaatag
      781 aggaaatacg gtaattgaat tttagtgaaa cttgcgacag ttttcctcat ttttttgtta
      841 ttccggcata cgagtgtggc atacgagtga ggtcatcttt tgtttcttcc gtttcttcat
      901 cgcttttgaa aaaaaatgtt gaagaaaact tcgtagccga tagccggata atgtttggca
      961 cggcgttcca acatatatca ttggaaattt tttaagattt ttgcgaaaaa tcacattctt
     1021 cacgatgaga acacgttatt gaaggaatat aaatcaagaa taacatatag ttatatttct
     1081 ctattacttt aacgttaaat atgagcaaat ttgagcattt tgattgcgat gaaaagcaga
     1141 atcgagtcaa ctgaaatccg ttcaaaaacc taagctcggt tcgcccgaaa gtcgattttt
     1201 ccaatagccg aacagcactg cttctttatt ttccaggtat ttttggggtg taagaattat
     1261 atcttctttg tttgggtatt tacgtttccg aattcccttt cttgatctgt cgctattttt
     1321 cagaaaccag aacagtttat cctttttcat gagaacctga atagctcaaa aacccgttta
     1381 atttcttttc atcatctcat ttgaaacttt cctagtcttc ttaatgattt ccaggctcct
     1441 cctacatttg tatctcaaga tgcatacact ttatttccgc tccaattcga atcatctact
     1501 tctctttttt cttcaacttt ctactttttt ccattctttt ctttcaaatg ttcgacgtct
     1561 tcaagtgact gctcttcatt ctcctcattt ttctcgaatt tcattcttgt ctcttttttg
     1621 ctcaaaaatg gaaaatgaaa gtgacgtgag atttggacgg cgggacacgg gggcagtaga
     1681 agcagcaaaa aggagagaaa gaggacacaa ataagaagaa cgaattcaaa aataagcgga
     1741 gaggagctat ttccgtcaat tctacctccc caatcttcat caattcggct caattgaatg
     1801 acgtcacagg gagataaacc gtttggatga gcgccgtcca tcactgacgc catcccgttt
     1861 gggacaagaa aagagaaaaa agagcacaaa gtttttggtg acggatcttg tcaatcatat
     1921 gaaagttgtt ctgattgatt gtcagttttt ttcctacttt tttggattct atccacttct
     1981 gaacttttga caagtttcaa actttctgaa tcattttcta tgcattttcc tggaattctt
     2041 tttatgtaaa atatgaaata gaatgttttt gaattcaagt tctgcttttt tcttcttttc
     2101 tgttctgttc gggcttgggt atgctttttt taaaaattat tttgcacatc gaccaataag
     2161 tgcgcaactt ataaaattaa tttatttttg ttaatttttg aaatacttgt attgctttaa
     2221 gtgatctgac ctcgccctga gctttccacg tagttatcaa atacaaatcc tccaagggta
     2281 acgtacctat attactgatc tttataataa ctttatcacc tgtccagttc cagaggaatt
     2341 ctgttaagct tataggcaca gaaggagtca tttctgctgg tttttaatga tccaaatctt
     2401 tatttcaagt aaaaaactga acacttgcga ataaaactat cagattaacc attcaccaaa
     2461 aatgtgtttg aatctaaaac tttctcagta ttccaaatat agaaataaat aaccacgaca
     2521 ttgctaaaat ctgtctgaat tgtgtactcc ttaccgtgaa gtaataatgg atataatgaa
     2581 tcgtttgaaa tgaatgatca gcacattttt ggtgaaagat cacaaataag gaataagcgt
     2641 cggaaaataa aacgattatt tcggatcaaa aatttgttga agatcatata cactcgagac
     2701 caagattatt ctagacaatt ttcaaattgg cttctttgtt tgcaaatctt cataataatc
     2761 tgtgaagttt ggaaatttga atttttaatc ttttttcata gattatagtt tttatttctt
     2821 tgcaaaacta tattaaaaac cgatgcattg ttttagggaa attaatgagc cttttgttca
     2881 acactaaaaa caataaaatt aaaattttgg cttcatcatt tgaccttttt taagttcgaa
     2941 aacttttctc gtatttctga accgccaatt ttttcacaca tctctagact tttggtgccc
     3001 gttccagaaa gttaagtaat tgctattcta agaaagttct caacattgtt ttttagttct
     3061 gattgaattc tgatgttcca ggaatatatt ttaaatttaa tattctttgc actatttcta
     3121 tactaactaa ataataaata gtctaagtat gttcaatgag aacaaaaaag ctctatctat
     3181 atttttctat cctatttcat ttatatcctt ttcattttga actcaccagc ttctcttctt
     3241 cttcttgtcc atatacttct taaaagtctc gaacttctct ctctcactct ccatcaattt
     3301 ctttatggct accgcttgcg ctctgccgct tccgaagaaa gaaggcgttg gcagagctct
     3361 caattctatt ttttctcgcc gtaggctatc gatttctcag cttctcttct ctcttcctca
     3421 ctccaccacc accacttcgg gcttcttctt ttcttcattt tcgtcttctt cttcatcctc
     3481 ttcttttttt ttcagttccc cctcactcct ccccttatat gcgcgtgcga gaggggtgca
     3541 aaagcagcgc gcatccgaga attgaaacga gaaaacggag acgcagcagc agttcgtcct
     3601 cagaaaaata gccagtaaaa agagaaaaag atagagagag acctatcgca ttttattttc
     3661 aaattgcaat tcctgcaatt catgtgtgcg tgtgcccatt ttcaattctt cccgttattt
     3721 ttcagaccat taccaacgtt ctttctagat cttgagacaa tcttccttct gctcaatcgt
     3781 tcgtcgtgaa gacggcatcg acgacgacga cgatcagcag taaaggatcg atccccggga
     3841 ttggccaaag gacccaaagg tatgtttcga atgatactaa cataacatag aacattttca
     3901 ggaggaccct tggctagcgt cgacggatcc aaaaaatggg atctcaccac caccaccacc
     3961 acggaatggc ctccatgacc ggaggacagc agatgggacg tgacctctac gacgacgacg
     4021 acaaggacct cgccactatg gtcgactcct cccgtcgtaa gtggaacaag accggacacg
     4081 ccgtccgtgc catcggacgt ctctcctccc tcgagaacgt ctacatcaag gccgacaagc
     4141 agaagaacgg aatcaaggta agtttaaaca tgattttact aactaactaa tctgatttaa
     4201 attttcaggc caacttccac atccgtcaca acatcgagga cggaggagtc cagctcgcct
     4261 accactacca gcagaacacc ccaatcggag acggaccagt cctcctccca gacaaccact
     4321 acctctccgt ccagtccaag ctctccaagg acccaaacga gaagcgtgac cacatggtcc
     4381 tccttgagtt cgtcaccgcc gccggaatca ccctcggaat ggacgagctc tacaagggag
     4441 gaaccggagg ttccatggtc tccaagggag aggagctctt caccggagtc gtcccaatcc
     4501 tcgtcgagct cgacggagac gtcaacggac acaagttctc cgtctctgga gaggtaagtt
     4561 taaacatgat tttactaact aactaatctg atttaaattt tcagggagag ggagacgcca
     4621 cctacggaaa gctcaccctc aagttcatct gcaccaccgg aaagctccca gtcccgtggc
     4681 cgaccctcgt caccaccctc acctacggag tccagtgctt ctcccgttac ccagaccaca
     4741 tgaagcagca cgacttcttc aagtccgcca tgccagaggg atacatccag gagcgtacca
     4801 tcttcttcaa ggacgacgga aactacaaga cccgtgccga ggtcaagttc gagggagaca
     4861 ccctcgtcaa ccgtatcgag ctcaagggaa tcgacttcaa ggaggacgga aacatcctcg
     4921 gacacaagct tgagtacaac ctcccagacc agctcaccga ggagcagatc gccgagttca
     4981 aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt caggaggcct
     5041 tctccctctt cgacaaggac ggagacggaa ccatcaccac caaggagctc ggaaccgtca
     5101 tgcgttccct cggacagaac ccaaccgagg ccgagctcca ggacatgatc aacgaggtcg
     5161 acgccgacgg agacggaacc atcgacttcc cagagttcct caccatgatg gcccgtaaga
     5221 tgaagtaccg tgacaccgag gaggagatcc gtgaggcctt cggagtcttc gacaaggacg
     5281 gaaacggata catctccgcc gccgagctcc gtcacgtcat gaccaacctc ggagagaagc
     5341 tcaccgacga ggaggtcgac gagatgatcc gtgaggccga catcgacgga gacggacagg
     5401 tcaactacga ggagttcgtc cagatgatga ccgccaaggg aggacgtgga ggatctggcc
     5461 caccggtcat gaccgctcca aagaagaaac gcaaagtacc gagctcagaa aaaatgacgg
     5521 cacctaaaaa gaaacgcaaa gtaccggtcg ccgagaagac agccccaaaa aagaagcgta
     5581 aagttccacg cgtcatggga gtaagtttaa acagttcggt actaactaac catacatatt
     5641 taaattttca gcgcgctgtc tccaagggag aggagctcat caaggagaac atgcacacca
     5701 agctctacat ggagggaacc gtcaacaacc accacttcaa gtgcacccac gaaggggagg
     5761 gcaagcccta cgagggcacc cagaccaaca ggattaaggt ggtggaggga ggccccctgc
     5821 cgttcgcatt cgacatcctg gccacctgct ttatgtacgg gagcaagacc ttcatcaacc
     5881 acacccaagg aatcccagac ttcttcaagc aatccttcgt aagtttaaac atgattttac
     5941 taactaacta atctgattta aattttcagc ctgagggctt cacatgggag agagtcacca
     6001 catacgaaga cgggggcgtg cttaccgtta cccaggacac cagcctccag gacggctgct
     6061 tgatctacaa cgtcaagctc agaggggtga acttcccatc caacggccct gtgatgcaga
     6121 agaaaacact cggctgggag gccagtaccg agaccctgta ccccgctgac ggcggcctgg
     6181 aaggcagatg cgacatggcc ctgaagctcg tgggcggggg ccacctgcac tgcaacctga
     6241 agaccacata cagatccaag aaacccgcta agaacctcaa gatgccagga gtaagtttaa
     6301 acatatatat actaactaac cctgattatt taaattttca ggtctacttt gtggaccgca
     6361 gactggaaag aatcaaggag gccgacaatg agacctacgt cgagcagcac gaggtggctg
     6421 tggccagata ctgcgacctc cctagcaaac tggggcacaa acttaatggc atggacgagc
     6481 tgtacaagta agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg
     6541 gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat
     6601 tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     6661 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     6721 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     6781 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     6841 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     6901 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     6961 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     7021 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     7081 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     7141 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     7201 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     7261 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     7321 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     7381 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7441 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7501 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7561 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     7621 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     7681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7741 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7801 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7861 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     8041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     8101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     8221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     8281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     8341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8641 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     9001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     9061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     9121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     9181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     9241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     9301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     9361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     9421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9481 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     9541 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9601 gaagagcgcc caatacgcaa accgcc
//
