LOCUS       sequence-404991-        5013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of WHaloCaMP1b-EGFP fusion in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  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 5013)
  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: pRSET-WHaloCaMP1b-EGFP
FEATURES             Location/Qualifiers
     source          1..5013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             163..180
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             745..1185
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1621..1623,1624..1626,1627..2337)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2414..2432)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2428..2475
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2572..3027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2659..2678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2869..2890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3053..3157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3158..3226,3227..4018)
                     /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(3376..3395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4189..4777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4678..4697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4931..4948
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgctgcagaa cgagcttgct
      121 cttaagttgg ctggacttga tattaacaag actggaggtt ctcatcatca ccaccaccat
      181 ggatccatgg cagaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg
      241 ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg
      301 cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc
      361 catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt
      421 tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa
      481 gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc
      541 aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg
      601 gacgaatggc cagaatttgc ccgcgagacc ttccagtggt tccgcaccgc ccgtcgtaaa
      661 tggcagaaaa caggccatgc ggttcgtgct atcggtcgtt tgtcgagcgg aggctctggc
      721 ggttccggcg gttcaggtgg atcagatcaa ttaacagagg aacagattgc ggagtttaag
      781 gaagcgttct ctttatttga taaggatggc gacggtacaa tcactactaa agaattggga
      841 acagtcatgc gctcattggg gcaaaatccg acagaggctg aattgcagga catgattaac
      901 gaggtagacg ccgatgggaa cgggactatc gactttccgg aatttcttac tatgatggca
      961 cgcaaaatga aagataccga ttctgaagaa gagatccgtg aagccttccg tgtttttgat
     1021 aaagatggga acggctacat cagtgctgct gagttacgcc atgtgatgac aaatctgggg
     1081 gaaaaactta ccgacgaaga agtagacgaa atgattcgcg aggcggatat tgacggggat
     1141 ggacaagtaa actacgagga atttgtgcag atgatgaccg ccaaggtcga ccgcaagctg
     1201 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
     1261 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     1321 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     1381 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     1441 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
     1501 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
     1561 atcggcagcg agatcgcgcg ctggctgtcg acgctcgaga tttccggcgg cggaagcggc
     1621 atggtctcta agggggagga actctttact ggtgtggtgc caatacttgt agagctcgat
     1681 ggggacgtta atggacacaa attttctgtc tcaggtgagg gtgagggaga tgcaacttat
     1741 ggaaaactga cattgaaatt catctgcaca acaggcaaat tgcctgtccc ctggcctacc
     1801 ctggtaacca ccctcactta tggcgttcag tgcttctccc ggtatcctga tcacatgaaa
     1861 cagcatgatt tcttcaaatc agcaatgccc gaaggttatg ttcaagagcg gaccatcttt
     1921 ttcaaggacg atggaaacta taaaaccaga gctgaggtta agttcgaggg agatactttg
     1981 gtgaatcgga tcgaactgaa ggggatagac ttcaaagagg atgggaacat tttggggcat
     2041 aaactcgaat ataactacaa ctcccacaac gtgtatatca tggctgacaa acagaagaac
     2101 gggatcaaag taaattttaa gatacgacac aatatagagg acggtagtgt ccaacttgct
     2161 gatcactacc agcagaatac acccatcgga gacggacccg ttctcttgcc cgataatcac
     2221 tatctctcta cacaatccgc tctgtctaaa gatccaaacg aaaagcggga ccacatggta
     2281 ctgcttgagt tcgttacagc cgctggtatc accttgggca tggatgagtt gtataaataa
     2341 gaattcgaag cttgatccgg ctgctaacgg tctcgactat acgcccgaaa ggaagctgag
     2401 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2461 ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat agcgaagagg
     2521 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2581 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2641 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2701 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2761 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2821 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2881 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2941 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3001 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3061 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaacaacc
     3121 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3181 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3241 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3301 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3361 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3421 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3481 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3541 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3601 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3661 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3721 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3781 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3841 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3901 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3961 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4021 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4081 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4141 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4201 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4261 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4321 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4381 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4441 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4501 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4561 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4621 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4681 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4741 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4801 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4861 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4921 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4981 tctccccgcg cgttggccga ttcattaatg cag
//
