LOCUS       sequence-372212-        7511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent-based opioid sensor for the mu opioid receptor in 
            AAV viral vector under a CAG promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7511)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7511)
  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            "unpublished"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-red-SPOTIT2
FEATURES             Location/Qualifiers
     source          1..7511
                     /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"
     CDS             2348..2404
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWIKAGHAVRAIGRLSS"
     primer_bind     3100..3119
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3977..4565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4030..4050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4448..4459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4608..4729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4619..4638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4673..4692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4891..5020
                     /label=AAV2 ITR
     rep_origin      complement(5257..5845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5337..5356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6016..6807,6808..6876))
                     /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     6639..6658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6877..6981)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7007..7462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7144..7165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7356..7375
                     /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 accttcggcg aatcacacgg 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 ccatggtcga
     2341 ctcatcacgt cgtaagtgga ttaaggcagg tcacgcagtc agagctatag gtcggctgag
     2401 ctcacccgtg gtttccgagc ggatgtaccc cgaggacggc gtcctgaaga gcgagatcaa
     2461 gaaggggctg aggctgaagg acggcggcca ctacgccgcc gaggtcaaga ccacctacaa
     2521 ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc gacataaagt tggacatcgt
     2581 gtcccacaac gaggactaca ccatcgtgga acagtgcgaa cgcgccgagg gccgccaccc
     2641 caccggcggc agggacgagc tgtacaaggg aggtacagga gggagtctgg tgagcaaggg
     2701 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2761 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggc
     2821 ctttcagacc gctaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     2881 cctgtcccct cagttcacat acggctccaa ggcctacatt aagcacccag ccgacatccc
     2941 cgactacttt aagctgtcct tccccgaggg cttcaggtgg gagcgcgtga tgaacttcga
     3001 ggacggcggc attattcacg ttaaccagga ctcctccctg caggacggcg tattcatcta
     3061 caaggtgaag ctgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     3121 catgggctgg gaggctacgc gtgaccaact gactgaagag cagatcgcag aatttaaaga
     3181 ggctttctcc ctatttgaca aggacgggga tgggacgata acaaccaagg agctggggac
     3241 ggtgttgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga
     3301 agtagatgcc gacggtgacg gcacattcga cttccctgag ttcctgacga tgatggcaag
     3361 aagaatgaat gacacagaca gtgaagtgga aattagagaa gcgttccgcg tgtttgataa
     3421 tgacggcaat ggctacatcg gcgcagcaga gcttcgccac gtgatgacag accttggaga
     3481 gaagttaaca gatgaggagg ttgatgaaat gatcagggta gcagacatcg atggggatgg
     3541 tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aaggatatca tggctcaagt
     3601 ccaacttgta gagtctggag gaggtctcgt gcggcccggc ggtagcctgc gactctcctg
     3661 tgttgattca gagaggacga gctaccctat ggggtggttt agacgggcac ccggaaagga
     3721 gcgcgaattc gttgcaagta tcacgtggag tggcattgac ccgacttatg ctgatagcgt
     3781 tgccgaccgg tttactacgt ccagagacgt tgcgaataac acgctgtacc tccagatgaa
     3841 ctctctcaag cacgaagata cagcggtcta ttactgtgct gcaagagcac ccgtcggtca
     3901 atcttcctct ccttatgatt atgactattg gggacagggg actcaggtaa ctgtgtccag
     3961 ttagaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4021 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4081 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4141 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4201 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4261 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4321 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4381 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4441 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4501 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4561 cccgcatcga taccgtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     4621 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4681 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4741 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     4801 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     4861 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     4921 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4981 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca
     5041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     7021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     7381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     7441 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7501 cgatcggtgc g
//
