LOCUS       sequence-405018-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of WHaloCaMP1a-EGFP in mammalian cells from the CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  AUTHORS   Farrants H, Shuai Y, Lemon WC, Hernandez CM, Yang S, Patel R, Qiao 
            G, Frei MS, Grimm JB, Hanson TL, Tomaska F, Turner GC, Stringer C, 
            Keller PJ, Beyene AG, Chen Y, Liang Y, Lavis LD, Schreiter ER
  TITLE     A modular chemigenetic calcium indicator enables in vivo functional 
            imaging with near-infrared light.
  JOURNAL   bioRxiv. 2023 Jul 19:2023.07.18.549527. doi: 
            10.1101/2023.07.18.549527. Preprint.
  PUBMED    37503182
REFERENCE   2  (bases 1 to 8173)
  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: "bioRxiv. 
            2023 Jul 19:2023.07.18.549527. doi: 10.1101/2023.07.18.549527. 
            Preprint."
COMMENT     Sequence Label: pCAG-WHaloCaMP1a-EGFP
FEATURES             Location/Qualifiers
     source          1..8173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..664
                     /label=chicken beta-actin promoter
     intron          665..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1689..1708
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2497..2937
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(3304..3306,3307..3309,3310..4020)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          4069..4201
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4184..4203
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4210..4798
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4263..4283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4681..4692)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4825..4842)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4831..4942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5038..5058)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5038..5056)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5077..5093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5195..5214)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5260..5315
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5314..5333)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5676..5692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5700..5716
                     /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)."
     promoter        complement(join(5724..5730,5731..5748,5749..5754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5849..6044
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5895..6030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6050..6184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6087..6106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6141..6160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6252..6269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6423..7011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6503..6522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7182..7973,7974..8042))
                     /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     7805..7824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8043..8147)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggncgaggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1081 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1621 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcggtac
     1741 cgcgggcccg gcgcgccagc tggatatctc cggacccggg ctagcgcaat tgcggccgcc
     1801 accatgctgc agaacgagct tgctcttaag ttggctggac ttgatattaa caagactgga
     1861 ggttctgcag aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc
     1921 gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac
     1981 ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat
     2041 cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat
     2101 ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag
     2161 gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat
     2221 ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac
     2281 gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc
     2341 aagctgatca tcgatcagaa cgtttttatc gagtggacgc tgccgatggc cgtcgcgcgc
     2401 gcccgtcgta aatggcagaa aacaggccat gcggttcgtg ctatcggtcg tttgtcgagc
     2461 ggaggctctg gcggttccgg cggttcaggt ggatcagatc aattaacaga ggaacagatt
     2521 gcggagttta aggaagcgtt ctctttattt gataaggatg gcgacggtac aatcactact
     2581 aaagaattgg gaacagtcat gcgctcattg gggcaaaatc cgacagaggc tgaattgcag
     2641 gacatgatta acgaggtaga cgccgatggg aacgggacta tcgactttcc ggaatttctt
     2701 actatgatgg cacgcaaaat gaaagatacc gattctgaag aagagatccg tgaagccttc
     2761 cgtgtttttg ataaagatgg gaacggctac atcagtgctg ctgagttacg ccatgtgatg
     2821 acaaatctgg gggaaaaact taccgacgaa gaagtagacg aaatgattcg cgaggcggat
     2881 attgacgggg atggacaagt aaactacgag gaatttgtgc agatgatgac cgccaagtat
     2941 ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag
     3001 ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg
     3061 ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg
     3121 ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct
     3181 aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac
     3241 ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg cggcggaagc
     3301 ggcatggtct ctaaggggga ggaactcttt actggtgtgg tgccaatact tgtagagctc
     3361 gatggggacg ttaatggaca caaattttct gtctcaggtg agggtgaggg agatgcaact
     3421 tatggaaaac tgacattgaa attcatctgc acaacaggca aattgcctgt cccctggcct
     3481 accctggtaa ccaccctcac ttatggcgtt cagtgcttct cccggtatcc tgatcacatg
     3541 aaacagcatg atttcttcaa atcagcaatg cccgaaggtt atgttcaaga gcggaccatc
     3601 tttttcaagg acgatggaaa ctataaaacc agagctgagg ttaagttcga gggagatact
     3661 ttggtgaatc ggatcgaact gaaggggata gacttcaaag aggatgggaa cattttgggg
     3721 cataaactcg aatataacta caactcccac aacgtgtata tcatggctga caaacagaag
     3781 aacgggatca aagtaaattt taagatacga cacaatatag aggacggtag tgtccaactt
     3841 gctgatcact accagcagaa tacacccatc ggagacggac ccgttctctt gcccgataat
     3901 cactatctct ctacacaatc cgctctgtct aaagatccaa acgaaaagcg ggaccacatg
     3961 gtactgcttg agttcgttac agccgctggt atcaccttgg gcatggatga gttgtataaa
     4021 taaaccggtg ctcgaggtac cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag
     4081 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     4141 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     4201 ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4261 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4321 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4381 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4441 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4501 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4561 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4621 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4681 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4741 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4801 ggaattcgag ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4861 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4921 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4981 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct
     5041 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5101 ttgttatccg ctaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     5161 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     5221 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     5281 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     5341 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     5401 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     5461 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     5521 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     5581 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     5641 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5821 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     5881 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5941 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6001 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     6061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6181 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6241 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6421 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6601 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6841 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7321 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7441 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7621 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7921 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8041 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8101 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8161 aaagtgccac ctg
//
