LOCUS       sequence-156216-        6924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of hM4D(Gi) using the CD68 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  AUTHORS   Grace PM, Strand KA, Galer EL, Urban DJ, Wang X, Baratta MV, 
            Fabisiak TJ, Anderson ND, Cheng K, Greene LI, Berkelhammer D, Zhang 
            Y, Ellis AL, Yin HH, Campeau S, Rice KC, Roth BL, Maier SF, Watkins 
            LR
  TITLE     Morphine paradoxically prolongs neuropathic pain in rats by 
            amplifying spinal NLRP3 inflammasome activation.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jun 14;113(24):E3441-50. doi: 
            10.1073/pnas.1602070113. Epub 2016 May 31.
  PUBMED    27247388
REFERENCE   2  (bases 1 to 6924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1602070113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-06-14- 14"; volume: "113"; issue: "24"; pages: 
            "E3441-50"
COMMENT     Sequence Label: pAAV CD68-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      850..1438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1339..1358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1500..1640
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1511..1640
                     /label=AAV2 ITR
     regulatory      2351..2360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3809..3818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3815..4525
                     /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(3960..3978)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4228..4247
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4550..5138
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4603..4623)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5021..5032)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5170..5646
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5369..5388)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5686..5826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5686..5815
                     /label=AAV2 ITR
     rep_origin      5901..6356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5988..6007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6198..6219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6373..6392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6492..6514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6552..6570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6638..6742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6743..6811,6812..6924,1..679)
                     /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"
ORIGIN
        1 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
       61 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      121 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      181 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      241 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      301 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      361 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      421 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      481 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      541 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      601 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      661 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      721 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      961 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc
     1501 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     1561 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     1621 ctccatcact aggggttcct gcggccgcac gcgtgatatc aaactgcctg tttgggcttc
     1681 tcatttctta cctccccttc cctctcccac ctgctactgg gtgcatctct gctcccccct
     1741 tccccagcag atggttacct ttgggctgtt gctttcttgt caccatctga gttctcagac
     1801 gctggaaagc catgttctcg gctctgtgaa tgacaatgct gactggagtg ctgcccctct
     1861 gtaaagggct gggtgtggat ggtcacaagc ccctcacatg cctcagccaa gaggaagtag
     1921 tacaggggtc agcccagagg tccaggggaa aggagtggaa accgatttcc ccaccaaggg
     1981 aggggcctgt acctcagctg ttcccatagc tacttgccac aactgccaag caagtttcgc
     2041 tgagtttgac acatggatcc ctgtggatca actgccctag gactccgttt gcacccatgt
     2101 gacactgttg actttgccct gacgaagcag ggccaacagt cccctaactt aattacaaaa
     2161 actaatgact aagagagagg tggctagagc tgaggcccct gagtcaggct gtgggtggga
     2221 tcatctccag tacaggaagt gagactttca tttcctcctt tccaagagag ggctgaggga
     2281 gcagggttga gcaactggtg cagacagcct agctggactt tgggtgaggc ggttcagcca
     2341 tatcgtcgac gccaccatgg ccaacttcac acctgtcaat ggcagctcgg gcaatcagtc
     2401 cgtgcgcctg gtcacgtcat catcccacaa tcgctatgag acggtggaaa tggtcttcat
     2461 tgccacagtg acaggctccc tgagcctggt gactgtcgtg ggcaacatcc tggtgatgct
     2521 gtccatcaag gtcaacaggc agctgcagac agtcaacaac tacttcctct tcagcctggc
     2581 gtgtgctgat ctcatcatag gcgccttctc catgaacctc tacaccgtgt acatcatcaa
     2641 gggctactgg cccctgggcg ccgtggtctg cgacctgtgg ctggccctgg actgcgtggt
     2701 gagcaacgcc tccgtcatga accttctcat catcagcttt gaccgctact tctgcgtcac
     2761 caagcctctc acctaccctg cccggcgcac caccaagatg gcaggcctca tgattgctgc
     2821 tgcctgggta ctgtccttcg tgctctgggc gcctgccatc ttgttctggc agtttgtggt
     2881 gggtaagcgg acggtgcccg acaaccagtg cttcatccag ttcctgtcca acccagcagt
     2941 gacctttggc acagccattg ctggcttcta cctgcctgtg gtcatcatga cggtgctgta
     3001 catccacatc tccctggcca gtcgcagccg agtccacaag caccggcccg agggcccgaa
     3061 ggagaagaaa gccaagacgc tggccttcct caagagccca ctaatgaagc agagcgtcaa
     3121 gaagcccccg cccggggagg ccgcccggga ggagctgcgc aatggcaagc tggaggaggc
     3181 ccccccgcca gcgctgccac cgccaccgcg ccccgtggct gataaggaca cttccaatga
     3241 gtccagctca ggcagtgcca cccagaacac caaggaacgc ccagccacag agctgtccac
     3301 cacagaggcc accacgcccg ccatgcccgc ccctcccctg cagccgcggg ccctcaaccc
     3361 agcctccaga tggtccaaga tccagattgt gacgaagcag acaggcaatg agtgtgtgac
     3421 agccattgag attgtgcctg ccacgccggc tggcatgcgc cctgcggcca acgtggcccg
     3481 caagttcgcc agcatcgctc gcaaccaggt gcgcaagaag cggcagatgg cggcccggga
     3541 gcgcaaagtg acacgaacga tctttgccat tctgctggcc ttcatcctca cctggacgcc
     3601 ctacaacgtc atggtcctgg tgaacacctt ctgccagagc tgcatccctg acacggtgtg
     3661 gtccattggc tactggctct gctacgtcaa cagcaccatc aaccctgcct gctatgctct
     3721 gtgcaacgcc acctttaaaa agaccttccg gcacctgctg ctgtgccagt atcggaacat
     3781 cggcactgcc aggcgggatc caccggtcgc caccatggtg agcaagggcg aggaggataa
     3841 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     3901 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3961 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4021 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     4081 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4141 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4201 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4261 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     4321 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     4381 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     4441 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4501 cggcatggac gagctgtaca agtaagaatt cgatatcaag cttatcgata atcaacctct
     4561 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4621 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4681 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4741 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4801 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4861 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4921 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     4981 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5041 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5101 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     5161 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     5221 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     5281 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     5341 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     5401 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     5461 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     5521 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     5581 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     5641 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     5701 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     5761 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     5821 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5881 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     6121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6241 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     6361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6481 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6541 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6901 gatccttgag agttttcgcc ccga
//
