LOCUS       sequence-372214-        7004 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 7004)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7004)
  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            "unpublished"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..7004
                     /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"
     repeat_region   4384..4513
                     /label=AAV2 ITR
     rep_origin      complement(4750..5338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4830..4849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5509..6300,6301..6369))
                     /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     6132..6151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6370..6474)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6500..6955)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6637..6658)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6849..6868
                     /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 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4501 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4561 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4621 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4681 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4741 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4801 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4861 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4921 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4981 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5041 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5101 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5161 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5221 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5281 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5341 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5401 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5461 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5521 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5581 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5641 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5701 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5761 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5821 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5881 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5941 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6001 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6061 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6121 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6181 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6241 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6301 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6361 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6421 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6481 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6541 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6601 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6661 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6721 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6781 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6841 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6901 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     6961 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcg
//
