LOCUS       Exported                8331 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Co-expresses hM4Dnrxn and a red fluorescent label.
ACCESSION   .
VERSION     .
KEYWORDS    CAG:: mCherry-2a-hM4Dnrxn
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  AUTHORS   Stachniak TJ, Ghosh A, Sternson SM
  TITLE     Chemogenetic synaptic silencing of neural circuits localizes a 
            hypothalamus-->midbrain pathway for feeding behavior.
  JOURNAL   Neuron. 2014 May 21;82(4):797-808. doi: 
            10.1016/j.neuron.2014.04.008. Epub 2014 Apr 24.
  PUBMED    24768300
REFERENCE   2  (bases 1 to 8331)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8331
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        81..460
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          703..1719
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1643..1665
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1727..1746
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1819..1828
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2545..2598
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4036..4062
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4063..4089
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          4303..4435
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4418..4437
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4444..5032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4497..4517)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4915..4926)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5059..5076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5065..5176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5272..5292)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5272..5290)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5311..5327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5429..5448)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5494..5549
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5548..5567)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5910..5926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5934..5950
                     /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(5958..5988)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6003..6024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6083..6278
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6129..6264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6191..6210
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6284..6418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6321..6340)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6375..6394
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6486..6503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6657..7245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6737..6756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7416..8276)
                     /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     8039..8058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8277..50)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
       61 cgaaaagtgc cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac
      121 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      181 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      241 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      301 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      361 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      421 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
      481 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
      541 ttaattattt tgtgcagcga tgggggcggg ggggggtggg ggggcggggc gaggcggaga
      601 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      661 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      721 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      781 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      841 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      901 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
      961 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     1021 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     1081 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     1141 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
     1201 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
     1261 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     1321 gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     1381 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     1441 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     1501 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     1561 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     1621 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     1681 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     1741 attgtgctgt ctcatcattt tggcaaagaa ttcggtaccg cgggcccggg atccaccgga
     1801 agccacctgc atgcggccgc caccatggta tcaaaggggg aagaagataa catggcaatt
     1861 atcaaggagt tcatgcggtt taaggtgcac atggaaggca gcgtcaacgg gcacgagttt
     1921 gagatcgagg gagagggtga gggacgcccc tacgagggga cacagacagc caagctgaaa
     1981 gtcaccaaag gaggaccgtt gcctttcgcg tgggacatcc tttcaccgca gtttatgtac
     2041 gggtccaaag cgtatgtgaa gcatcccgcg gacattccag actatcttaa actgtcgttt
     2101 cccgagggct tcaaatggga gcgagtgatg aattttgagg atggaggggt agtgacagtg
     2161 acgcaagact cgtccctcca agacggcgag tttatctaca aggtgaaact cagaggcact
     2221 aatttcccct cggacggtcc agtcatgcag aaaaagacga tgggttggga ggcgtcgtcg
     2281 gagcggatgt atcccgaaga tggagcactc aaaggggaga tcaaacagag attgaagttg
     2341 aaagacggtg gacactacga cgccgaagtc aagacaacgt ataaggcgaa gaagccggtg
     2401 cagttgccgg gtgcctacaa tgtgaatatc aaactcgaca ttacaagcca taacgaggat
     2461 tatacgatcg tggaacagta cgaaagggcg gaggggcgac attcgaccgg gggaatggat
     2521 gagttgtata aggctagcag agcagaagga cgcgggagcc ttttgacgtg tggggatgtc
     2581 gaagagaatc cgggacctat ggcgaacttc acgcctgtca atgggagctc cggtaatcag
     2641 tcggtacggc tcgtgacgtc gtcctcgcac aatcgctacg aaacagtaga aatggtattc
     2701 atcgccacag taactggaag cctttcactt gtgactgtcg tgggaaacat tctggtcatg
     2761 ctctcgatta aggtcaacag gcaactccag accgtaaaca actatttctt gttctccctg
     2821 gcttgcgcgg atctgatcat tggggctttc tcgatgaacc tctatacagt atacatcatc
     2881 aaagggtact ggcctctcgg agccgtcgtg tgtgacctct ggcttgcact tgactgtgtg
     2941 gtgagcaatg cttcagtcat gaaccttctg attatctcat ttgatcgcta tttctgtgta
     3001 accaaaccct tgacctaccc cgcaaggaga acgaccaaaa tggcagggct gatgatcgcg
     3061 gctgcatggg tcctctcgtt cgtactttgg gctcctgcca ttctgttctg gcagtttgtc
     3121 gtgggtaaac gcacagtccc ggacaaccaa tgtttcatcc aattcctttc gaaccccgct
     3181 gtgactttcg ggactgcgat tgcgggattc taccttccgg tcgtcattat gaccgtgttg
     3241 tacattcaca tctcgcttgc gtcaagaagc cgcgtccaca agcacaggcc cgaggggcct
     3301 aaggagaaga aagccaagac attggctttt ctcaaatcac cgcttatgaa acagtccgtg
     3361 aaaaagcctc ctcccggtga agcagcgcgc gaggagctga ggaatggtaa actggaagag
     3421 gcgccacctc cggcgctgcc tcctccaccg cgacccgtgg cggataagga cacttccaac
     3481 gagtcgtcgt cggggagcgc gacccagaat acgaaagagc gacctgctac cgagttgagc
     3541 acaacagaag cgacgacgcc ggcaatgccg gcaccgcctc ttcagcccag ggcgctgaac
     3601 cccgcgtcgc gatggtcaaa gattcagatt gtgacgaaac aaacgggtaa tgagtgcgta
     3661 acagccatcg agatcgtacc cgcaacgccc gctggtatga ggccagccgc caacgtcgcc
     3721 agaaagttcg cgtcaatcgc gcggaatcag gtgcggaaga aaagacaaat ggccgcgaga
     3781 gagagaaagg tgacaagaac gattttcgcc atcctcctgg cgtttatctt gacgtggact
     3841 ccgtataatg tcatggtgtt ggtgaataca ttttgtcaat cgtgcatccc agacacggtc
     3901 tggtcgatcg ggtattggct ctgctacgtc aactccacta tcaatccggc atgctacgca
     3961 ttgtgtaacg ccacgttcaa gaaaacgttc cgccatctcc tgttgtgcca gtaccgaaat
     4021 atcgggacag cgagataccc atatgatgtc cccgattacg cctatcccta tgacgtcccg
     4081 gattacgctt acaagtatcg gaaccgggac gaaggctcgt atcatgtgga cgagtcacgg
     4141 aactatatct cgaactccgc ccagtcaaac ggagcggtgg tgaaggaaaa gcagccctca
     4201 tcggcgaagt cagcgaataa gaacaagaag aataaggaca aagagtacta tgtgtgaacc
     4261 ggtgctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat caaggttaca
     4321 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     4381 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     4441 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4501 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4561 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4621 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4681 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4741 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4801 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4861 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4921 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4981 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     5041 cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5101 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5161 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5221 ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt ccctttagtg
     5281 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5341 tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     5401 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     5461 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     5521 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     5581 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     5641 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     5701 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     5761 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     5821 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     5881 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6001 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6061 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     6121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6181 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6241 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     6301 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6361 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     6421 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6481 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6541 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6601 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6661 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6721 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6781 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6841 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6901 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6961 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7021 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7081 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7141 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7201 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7261 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7321 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7381 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7441 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7501 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7561 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7621 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7681 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7741 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7801 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7861 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7921 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7981 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8041 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8101 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8161 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8221 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8281 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag c
//
