LOCUS       sequence-373784-        7278 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of genetically encoded calcium indicator 
            SCW-HHL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7278)
  AUTHORS   Gohil K, Wu SY, Takahashi-Yamashiro K, Shen Y, Campbell RE
  TITLE     Biosensor Optimization Using a Forster Resonance Energy Transfer 
            Pair Based on mScarlet Red Fluorescent Protein and an 
            mScarlet-Derived Green Fluorescent Protein.
  JOURNAL   ACS Sens. 2023 Feb 24;8(2):587-597. doi: 10.1021/acssensors.2c01730.
            Epub 2023 Jan 24.
  PUBMED    36693235
REFERENCE   2  (bases 1 to 7278)
  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"; doi: 
            "10.1021/acssensors.2c01730"; journalName: "ACS Sens"; date: 
            "2023-02-24- 24"; volume: "8"; issue: "2"; pages: "587-597"
COMMENT     Sequence Label: pCDNA-SCW-HHL
FEATURES             Location/Qualifiers
     source          1..7278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2086..2142
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(2869..2886)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2875..3099
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3145..3573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3232..3251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3442..3463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3582..3602)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3587..3916
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3767..3902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3829..3848
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3983..4777
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4037..4056)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4647..4666
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5042..5061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5121..5137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5121..5137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5134..5156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5145..5161
                     /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(5169..5175,5176..5193,5194..5199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5214..5235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5352..5369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5523..6111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5603..5622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6282..7073,7074..7142))
                     /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     6905..6924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7143..7247)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagaatg gtgagcaagg gcgaggcagt gatcaaggag
      961 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     1021 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1081 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
     1141 gccttcatca agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc
     1201 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac
     1261 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     1321 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg
     1381 taccccgagg acggcgtgct gaagggcgac attcatatgg ccctgcgcct gaaggacggc
     1441 ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
     1501 ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg
     1561 gtggaacagt acgaacgctc cgagggccgc caccacctag gcggcgcatg cctgccggac
     1621 caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
     1681 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
     1741 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
     1801 atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa tgaaatacag ggacacggaa
     1861 gaagaaatta gagaagcgtt cggtgtgttt gataaggatg gcaatggcta catcagtgca
     1921 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
     1981 gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta
     2041 caaatgatga cagcgaaggg tggcggaggt tctgtcgact catcacgtcg taagtggaat
     2101 aagacaggtc acgcagtcag agctataggt cggctgagct cactagaggg cgaggcattg
     2161 atcaaggagt acatgcggtt caaggtgcac atggagggct ccatggacgg ccacgagttc
     2221 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccataccgc caagctgaag
     2281 gtgaccaagg gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcgggtac
     2341 ggctccaggg ccttcatcaa gcaccccgcc gacatccccg actactataa gcagtccttc
     2401 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg
     2461 acccaggaca cctccctgga ggacggcacc ctgatccaca aggtgaagct ccgcggcacc
     2521 aacttccctc ctgacggccc cgtaatgcag aggaagacaa tgggctggga agcgtccacc
     2581 gagcggttgt accccgagga cggcgtgctg aagggcgaca ttcatatggc cctgcgcctg
     2641 aaggacggcg gccgctacct ggcggactgc aagaccacct acaaggccaa gaagcccgtg
     2701 cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac
     2761 tacaccgtgg tggaacagta cgaacgctca gagggccgcc accacctagg cggcatggac
     2821 gagctgtaca agtaagaatt cgaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc
     2881 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2941 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3001 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3061 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3121 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3181 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3241 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3301 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3361 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3421 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3481 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3541 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3601 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3661 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3721 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3781 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3841 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3901 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3961 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4021 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4081 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4141 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4201 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4261 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4321 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4381 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4441 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4501 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4561 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4621 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4681 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4741 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4801 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4861 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4921 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5101 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5161 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5221 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5341 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5401 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5461 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5701 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5881 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5941 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6001 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6061 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6121 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6181 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6241 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6301 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6361 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6421 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6481 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6541 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6601 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6661 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6721 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6781 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6841 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6901 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6961 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7021 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7081 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7141 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7201 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7261 aaagtgccac ctgacgtc
//
