LOCUS       Exported                6165 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  FLiCRE reporter gene for neuronal expression.
ACCESSION   .
VERSION     .
KEYWORDS    TRE:mCherry-p2a- bReaChES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  AUTHORS   Kim CK, Sanchez MI, Hoerbelt P, Fenno LE, Malenka RC, Deisseroth K, 
            Ting AY
  TITLE     A Molecular Calcium Integrator Reveals a Striatal Cell Type Driving 
            Aversion.
  JOURNAL   Cell. 2020 Dec 23;183(7):2003-2019.e16. doi: 
            10.1016/j.cell.2020.11.015. Epub 2020 Dec 11.
  PUBMED    33308478
REFERENCE   2  (bases 1 to 6165)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6165
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    142..160
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    214..232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    249..267
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    285..303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        315..353
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     350..374
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      402..411
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             408..1115
                     /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"
     primer_bind     complement(553..571)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     821..840
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1125..1181
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2178..2204
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2223..2811
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2276..2296)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2694..2705)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2843..3319
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3042..3061)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3359..3499
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3359..3488
                     /label=AAV2 ITR
     rep_origin      3574..4029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3661..3680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3871..3892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4046..4065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4165..4187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4225..4243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4311..4415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4416..5276
                     /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(4634..4653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5447..6035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5936..5955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6097..61
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
       61 tgcggccgca cgcgtccttc aggtccgagg ttctagacga gtttactccc tatcagtgat
      121 agagaacgat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
      181 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      241 cgagtttatc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
      301 agaacgtatg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
      361 tgaaccgtca gatcgcaaag ggcgaattcg atccaccggt cgccaccatg gtgagcaagg
      421 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      481 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      541 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      601 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      661 ccgactactt gaagctgtcc ttccccgagg gattcaagtg ggagcgcgtg atgaacttcg
      721 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
      781 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
      841 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
      901 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
      961 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     1021 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     1081 gccactccac cggcggcatg gacgagctgt acaagggctc cggagccacg aacttctctc
     1141 tgttaaagca agcaggagac gtggaagaaa accccggtcc catggactat ggcggcgctt
     1201 tgtctgccgt cggactgttc cagaccagct acaccctgga gaacaacggc agcgtgatct
     1261 gcatccccaa caacggccag tgcttctgcc tggcctggct gaagagtaac ggcaccaacg
     1321 ccgagaagct ggccgccaac atcctgcagt gggtggtgtt tgcgctgagc gtggcgtgcc
     1381 tgggctggta tgcgtatcag gcgtggcgcg cgacctgcgg ctgggaagaa gtgtatgtgg
     1441 cgctgattga aatgatgaaa agcattattg aagcgtttca tgaatttgat agcccggcga
     1501 ccctgtggct gagcagcggc aacggcgtgg tgtggatgcg ctatggctca tggctgctga
     1561 cctgccccgt gattctgatt catctgagca acctgaccgg cctgaaagat gattatagca
     1621 aacgcaccat gggcctgctg gtgagcgacg tgggctgcat tgtgtggggc gcgaccagcg
     1681 cgatgtgcac cggctggacc aaaattctgt tttttctgat tagcctgagc tatggcatgt
     1741 atacctattt tcatgcggcc aaagtgtata ttgaagcgtt tcataccgtg ccgaaaggcc
     1801 tgtgcagaca gctggtgaga gccatggcct ggctgttctt cgtgagctgg ggcatgttcc
     1861 ccgtgctgtt cctgctgggc cccgagggct tcggccatat tagcccgtat ggcagcgcga
     1921 ttggccatag cattctggat ctgattgcga agaacatgtg gggcgtgctg ggcaactatc
     1981 tgcgcgtgaa aattcatgaa catattctgc tgtatggcga tattcgcaaa aaacagaaaa
     2041 ttaccattgc gggccaggaa atggaagtgg aaaccctggt ggcggaagaa gaagatgcgg
     2101 ccgccaagag caggatcacc agcgagggcg agtacatccc cctggaccag atcgacatca
     2161 acgtgggcgc gcccgggtac ccatacgatg tgccagatta cgcctaatag aagcttatcg
     2221 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2281 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2341 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2401 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2461 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2521 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2581 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2641 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2701 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2761 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     2821 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     2881 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2941 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     3001 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     3061 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     3121 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     3181 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     3241 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     3301 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     3361 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3421 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3481 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3541 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3601 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3661 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3721 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3781 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3841 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3901 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3961 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4021 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4081 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4141 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4201 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4261 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4381 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4441 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4501 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4561 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4621 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4681 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4741 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4801 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4861 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4921 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4981 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5041 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5101 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5161 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5221 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5281 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5341 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6061 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     6121 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccg
//
