LOCUS       sequence-405001-        4224 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of WHaloCaMP1a in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  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 4224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-WHaloCaMP1a
FEATURES             Location/Qualifiers
     source          1..4224
                     /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             103..120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             754..1194
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1625..1643)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1639..1686
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1783..2238
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1870..1889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2080..2101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2264..2368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2369..2437,2438..3229)
                     /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(2587..2606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3400..3988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3889..3908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4142..4159
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca ccaccaccat
      121 ggagcagaaa tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag
      181 cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt
      241 aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc
      301 tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc
      361 ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc
      421 gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca
      481 gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa
      541 tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag
      601 ctgatcatcg atcagaacgt ttttatcgag tggacgctgc cgatggccgt cgcgcgcgcc
      661 cgtcgtaaat ggcagaaaac aggccatgcg gttcgtgcta tcggtcgttt gtcgagcgga
      721 ggctctggcg gttccggcgg ttcaggtgga tcagatcaat taacagagga acagattgcg
      781 gagtttaagg aagcgttctc tttatttgat aaggatggcg acggtacaat cactactaaa
      841 gaattgggaa cagtcatgcg ctcattgggg caaaatccga cagaggctga attgcaggac
      901 atgattaacg aggtagacgc cgatgggaac gggactatcg actttccgga atttcttact
      961 atgatggcac gcaaaatgaa agataccgat tctgaagaag agatccgtga agccttccgt
     1021 gtttttgata aagatgggaa cggctacatc agtgctgctg agttacgcca tgtgatgaca
     1081 aatctggggg aaaaacttac cgacgaagaa gtagacgaaa tgattcgcga ggcggatatt
     1141 gacggggatg gacaagtaaa ctacgaggaa tttgtgcaga tgatgaccgc caagtatctg
     1201 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     1261 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     1321 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     1381 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
     1441 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
     1501 atcggcagcg agatcgcgcg ctggctgtcg acgctcgaga tttccggcta agaattcgaa
     1561 gcttgatccg gcttctaacg gtctcgacta tacgcccgaa aggaagctga gttggctgct
     1621 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1681 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     1741 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     1801 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1861 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1921 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1981 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2041 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2101 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2161 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2221 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2281 tttatttttc taaatacatt caaatatgta tccgctcatg agacaacaac cctgataaat
     2341 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2401 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2461 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2521 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2581 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2641 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2701 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2761 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2821 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2881 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2941 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3001 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3061 taaatctgga gctggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3121 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3181 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3241 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3301 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3721 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3781 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3841 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3901 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3961 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4021 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4081 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4141 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4201 gcgttggccg attcattaat gcag
//
