LOCUS       sequence-377870-        6982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent-based opioid sensor for the mu opioid receptor 
            with reduced Gai coupling in AAV viral vector under a CAG promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6982)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6982)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: AAV-CAG-M-SPOTIT2(R279F)
FEATURES             Location/Qualifiers
     source          1..6982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   23..152
                     /label=AAV2 ITR
     promoter        463..740
                     /label=chicken beta-actin promoter
     primer_bind     1019..1041
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1103..1122
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2551..2572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2678..2699)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2939..2958)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3470..4058
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3523..3543)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3941..3952)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4101..4222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4112..4131)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4166..4185
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4728..5316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4808..4827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5487..6278,6279..6347))
                     /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     6110..6129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6348..6452)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6478..6933)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6615..6636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6827..6846
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
       61 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      121 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
      181 cttatctacg tagccatgct ctaggaagag taccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      481 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      541 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
      601 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      661 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      721 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      781 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      841 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      901 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      961 gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1021 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1081 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     1141 aaagaattgg atccccacca tgagacccca gatcctgctg ctcctggccc tgctgaccct
     1201 aggcctggct gactacaagg acgatgatga cgccatggac tcttccgctg ctcccaccaa
     1261 tgcctcaaat tgtacagacg ccttggccta tagctcttgc tctcccgcac catctccggg
     1321 aagctgggtt aacctcagtc atctcgacgg gaacctgtcc gacccttgtg gccctaatag
     1381 gaccgacctc ggtggtcgag atagcctgtg cccccctact ggaagcccca gcatgatcac
     1441 agctatcaca ataatggctc tctatagtat cgtctgcgtc gtgggactct ttggcaattt
     1501 tctcgtcatg tacgttatcg tcagatacac aaagatgaaa accgcaacca atatttacat
     1561 ctttaatctg gccctggcgg atgcccttgc aaccagtaca ctgccgtttc aatccgtgaa
     1621 ttatctgatg ggcacatggc ccttcggaac cattctgtgc aaaatcgtaa tctcaattga
     1681 ttattataac atgttcacct ccatttttac cttgtgtaca atgtcagtag accgctatat
     1741 cgccgtatgc caccctgtca aagctcttga tttcaggacc cctaggaatg ccaagatcat
     1801 caatgtgtgc aattggattc tgtctagcgc tatcggcttg ccggtgatgt tcatggccac
     1861 taccaagtac agacaaggtt caatcgactg taccctcaca ttttcacatc ccacgtggta
     1921 ttgggagaac ttgctgaaga tctgtgtgtt catcttcgcc ttcatcatgc cagtactgat
     1981 aatcaccgtt tgttatggtc ttatgatcct gcgcctgaag agcgtgcgca tgctcagcgg
     2041 ctccaaagaa aaggaccgga accttcggtt catcacacgg atggtcctgg tggttgtggc
     2101 tgtgttcatt gtttgctgga cccctattca tatttatgtt ataattaagg ccctggtgac
     2161 catcccggaa accacattcc agaccgtttc ctggcacttt tgtatagcat tgggatatac
     2221 caacagttgc cttaatcccg ttctgtacgc gtttctggac gagaatttca agcgatgctt
     2281 cagggagttc tgcttccccc tcaagatgcg catggagcgg cagggcgcgc cctacaacag
     2341 ccacaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttccacat
     2401 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc
     2461 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact
     2521 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2581 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag
     2641 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2701 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct
     2761 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2821 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2881 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga
     2941 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3001 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3061 gtacaacctt aaggaagata tcatggctca agtccaactt gtagagtctg gaggaggtct
     3121 cgtgcggccc ggcggtagcc tgcgactctc ctgtgttgat tcagagagga cgagctaccc
     3181 tatggggtgg tttagacggg cacccggaaa ggagcgcgaa ttcgttgcaa gtatcacgtg
     3241 gagtggcatt gacccgactt atgctgatag cgttgccgac cggtttacta cgtccagaga
     3301 cgttgcgaat aacacgctgt acctccagat gaactctctc aagcacgaag atacagcggt
     3361 ctattactgt gctgcaagag cacccgtcgg tcaatcttcc tctccttatg attatgacta
     3421 ttggggacag gggactcagg taactgtgtc cagttagaag cttatcgata atcaacctct
     3481 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3541 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3601 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3661 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3721 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3781 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3841 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3901 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3961 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4021 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg
     4081 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4141 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4201 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4261 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     4321 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     4381 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4441 aggccgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa
     4501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6361 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6421 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     6481 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6541 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6601 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6661 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6721 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6781 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6841 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6901 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     6961 gttgggaagg gcgatcggtg cg
//
