LOCUS       7_fabp10a_GcAMP6        8558 bp    DNA     circular SYN 02-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8558)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 14, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..62
                     /label=T7 primer
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        70..2251
                     /label=fabp10a
     primer_bind     2050..2069
                     
     regulatory      2267..2276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2273..2281,2282..2299,2300..2302,2303..2335,2336..2338,
                     2339..2362,2363..2389,2390..2446,2447..2452,2453..2722,
                     2723..2740,2741..3175,3176..3181,3182..3625)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(2629..2648)
                     /label=GCaMP7f_Seq_RP p24
     primer_bind     2629..2648
                     /label=GCaMP7f_Ver_FP p25
     polyA_signal    3923..4057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3959..3980)
                     /label=Sv40pA_Seq_RP2 p41 box2
     protein_bind    complement(4068..4101)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    4109..4156
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    4180..4387
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4411..5121)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     promoter        complement(5127..5576)
                     /label=cryaa
     protein_bind    complement(5687..5720)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        complement(5739..5757)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5790..5806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5948..6403
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6429..6533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6534..6602,6603..7394)
                     /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"
     rep_origin      7565..8153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8441..8462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8477..8482,8483..8500,8501..8507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8515..8531
                     /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     8539..8555
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gattacgcca agcgcgttag ggataacagg gtaatgcgcg cgtaatacga ctcactatag
       61 ggcgaattgg agctctgcag aagagcatct ctgaacacac aacacgtcca accttgaggc
      121 agatgggcta cagcagcaga agaccacacc gggtgccgct cctgtcagct aagaacagga
      181 aactgaggct acaattcaca cagactcacc aaaactggac aatatattat tagcccccct
      241 tagaattttt catttgataa tatttttctt ctggcgaaag cctcatttgt tttatatatt
      301 atagaataaa attagttttt aatagttttt atgccatttt aaggtcaata ttattagccc
      361 ctttaagcta ttttttttcg atagtctaca gaacaaacca tcggtataca atgacttgcc
      421 taattaccct aacctgccta gttaccctaa ttaacctagt taagccttta aatgtcactt
      481 taagctgtat agaagtgtct tgaagaatat ctagtctaat attattgact gtcatcatgg
      541 caagataaaa taaatcagtt attaaaacta ttatgattag aaatgtgctg aaacaatctg
      601 ctctccgata aacagaaatt gaacaaaata aacagggggg ctaataaatt taaggggtta
      661 aataattctg attgcaaaaa aaatgatgtc tgcaaaactg ttgcaaataa tttatttgtg
      721 ttgattttaa gcaaacaaat taaatttaat aaaatacaac ttaatctgtt tgtttaaatt
      781 cagccctaat aaattgttta cagccactta acgtaaaaaa attgagtaaa tccaaggaat
      841 catctctgaa taattttttc agtgtatata tatatatata tattcttaca aaacaactca
      901 tttactttag ttaattttca ggggcaaaaa ctaaagtaat cgacgttgct tgaataaaaa
      961 gtgtaattaa gggaatgagg taacatttaa ccatgtgtca atgcagttta aatatgccag
     1021 ttagtggtat atgtttaaat ggtaagctat tcaaaacttt aaactaactt aaccagcctt
     1081 ttgttgtcag actgaacaga ctttccatct gcattattag agactaatct ttggctggat
     1141 gaatgattca tctgctgata tttcagaata gacagattga ggctgtttct aatatgatta
     1201 tgcaacctga gggtgattat ttgaagcaaa ctccacagac cagcaggtca ttgaccgtcg
     1261 tgtgttcaaa cagagcagaa acatttgcaa aactggtctg acaggagaat ccagtccagc
     1321 acaacacata tgctgagcaa actgaatcaa tcctgcaggt caactctcgt gctttaagtt
     1381 tattaaagat tattttattt atttattatt ttatttatct atttatttat ttagttgttt
     1441 atttattcct gcagatcatg ccttgtgcct ttttacattt aatttaattt ttaatttaat
     1501 ttccttttat ttttttttat ttttttattt tattttattt tacagtctga catatggacc
     1561 aataaatctc tcagatcatg tctcatgcat ttaatttaat ttttttattt attcattttt
     1621 atttatttat tttattattt attttatttt aatatttatt ttattttatt ttattttatt
     1681 ttacagtctg acatactgga ccaataaatc tctcagatca tgtctcatgc attttacttt
     1741 tattttatta gaattagaaa gatcaaagga acaactttta aaatattaat tctgtatcaa
     1801 aatctctttt gatacattta attgatttaa aaaagcagtt cacccaagaa acatttcctc
     1861 acagtcgaat ggttgtaaac ttttatgaat tactttcaca gaaaaagatt tttggaagaa
     1921 tattggaaaa aaagcagcca ttgacttcca tagtaacaac aaaaaatact atggaagtca
     1981 atggctgttt tttcaccatt cggtatcttc attctggagc agaatttttt gggtgatctg
     2041 tccctttaag tcgtcaaatc ctggtgcaat attccacatg cactgacgag tctttatttt
     2101 tggtctgcta ctgctgtgtg tgtgagggca tttacctcat cttctgctgg agttgatgaa
     2161 cggtgggttg ttcaaacagc agcaggtcat tgactgaact cctctcgata taaaagctgc
     2221 agatctgaag ctgaccttca ctttgtgttg atcgctagcg gatctcgcca ccatgggttc
     2281 tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga
     2341 tctgtacgac gatgacgata aggatctcgc caccatggtc gactcatcac gtcgtaagtg
     2401 gaataagaca ggtcacgcag tcagagctat aggtcggctg agctcactcg agaacgtcta
     2461 tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttccacatcc gccacaacat
     2521 cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg
     2581 ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc
     2641 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2701 cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagca agggcgagga
     2761 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2821 gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt
     2881 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2941 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3001 cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3061 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3121 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacctgcc
     3181 ggaccaactg actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa
     3241 ggacggggat gggacaataa caaccaagga gctggggacg gtgatgcggt ctctggggca
     3301 gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg
     3361 cacaatcgac ttccctgagt tcctgacaat gatggcaaga aaaatgaaat acagggacac
     3421 ggaagaagaa attagagaag cgttcggtgt gtttgataag gatggcaatg gctacatcag
     3481 tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt
     3541 tgatgaaatg atcagggaag cagacatcga tggggatggt caggtaaact acgaagagtt
     3601 tgtacaaatg atgacagcga agtgagcggc cgccgaaatg accgaccaag cgacgcccaa
     3661 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3721 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3781 cgcccacccc ccggatctaa gctctagata agtaatgatc ataatcagcc atatcacatc
     3841 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3901 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3961 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4021 gtccaaactc atcaatgtat cttatcatgt ctggatccgg gggtaccgaa gttcctatac
     4081 tttctagaga ataggaactt cgcgtcgaga agttcctatt ccgaagttcc tattctctag
     4141 aaagtatagg aacttcgggc ccgcggtggc ggcgatctct ccccagcatg cctgctattg
     4201 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     4261 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     4321 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     4381 ggcacagtcg aggctgatca gcgggccgct ttacttgtac agctcgtcca tgccgccggt
     4441 ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct cgttgtggga
     4501 ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc
     4561 cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct tcagcctctg
     4621 cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca
     4681 gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt
     4741 caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc
     4801 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa
     4861 gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg
     4921 ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca gcttggcggt
     4981 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     5041 cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg ccatgttatc
     5101 ctcctcgccc ttgctcacca tgttgtcaga cctggtaact ccttactgta acttattgca
     5161 cctgatgaac aaaagatgtg aactgaacgc aagtaggagg ctaatagagg ctagtacagt
     5221 acaaagatcc ccaaatgatg tgtgccggcc ttagtggagg cagacccctg gttatatacc
     5281 tgtgctgcag aaagaggggc ttacaaatgg atctctctgg aatggcatta tctcatggtg
     5341 ggaacagtgt tgtcagcaga atcagcgggg tggactctga cctgagaatg acagtctagc
     5401 actgaatggc tcagacaata ctcttgtggc caactatcac cggaaaataa gtgcctgcca
     5461 aaacaaacca gaccagatga agctacatgg ctgtagaacg tcagtattta atgtgttgtg
     5521 gccacttggt tcaccagatt taagagctaa tttttttaaa tatcacagat aatcaatcgc
     5581 tttttttttt tcttgactga aaagcttaga ctggaaaatt ctttgaaaaa gtttgcacta
     5641 cagaaaataa atacataaca tcctgcactg aatgcacact attaatgaag ttcctatact
     5701 ttctagagaa taggaacttc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     5761 cgctagggat aacagggtaa tacgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5821 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5881 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5941 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6001 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6061 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6121 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6181 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6241 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6301 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6361 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     6421 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6481 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6541 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6601 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6661 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6721 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6781 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6841 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6901 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6961 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7021 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7081 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7141 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7201 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7261 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7321 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7381 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7441 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7501 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7561 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7621 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7681 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7741 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7801 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7861 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7921 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7981 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8041 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8101 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8161 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8221 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8281 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8341 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8401 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8461 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8521 gcggataaca atttcacaca ggaaacagct atgaccat
//
