LOCUS       sequence-377895-        7905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent-based opioid sensor for the kappa 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 7905)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7905)
  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            "unpublished"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-K-red-SPOTIT(R271F)
FEATURES             Location/Qualifiers
     source          1..7905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     568..587
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1865..1921
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWIKAGHAVRAIGRLSS"
     primer_bind     2617..2636
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3494..4082
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3547..3567)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3965..3976)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4125..4246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4136..4155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4190..4209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4625..5213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4705..4724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5384..6175,6176..6244))
                     /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     6007..6026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6245..6349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6375..6830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6512..6533)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6724..6743
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6902..7031
                     /label=AAV2 ITR
ORIGIN
        1 ggatacggga gcggggtggg tctggggggc ggggaggggg gtagaaggga cggggccggg
       61 gggggcgggg gggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      121 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      181 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      241 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      301 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      361 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
      421 ggggagcggc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
      481 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
      541 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
      601 ttgggcaaag aattggatcc ccaccatgaa gacgattatc gccctgagct acatcttctg
      661 cctggtgttc gccgactaca aggacgatga tgacgccatg gacagcccca ttcaaatctt
      721 ccggggcgag cccggaccaa cctgcgctcc aagcgcttgc ctgccaccga attctagtgc
      781 atggttccca gggtgggcag agcccgatag caatggctca gccgggtccg aggatgcgca
      841 gcttgaaccc gctcacatct cacccgctat ccctgtcatt attacggcag tctactcagt
      901 tgttttcgtt gtgggcctgg ttggaaatag cctcgtgatg tttgtgatta tcaggtatac
      961 taaaatgaag accgccacga atatctacat ctttaacctg gctctggctg acgccttggt
     1021 gaccaccact atgccatttc agtccaccgt ctatcttatg aactcttggc cattcggcga
     1081 cgttctgtgc aagattgtca tttccattga ctattataat atgtttacat caatctttac
     1141 gcttacaatg atgagcgtgg accgatacat tgctgtttgc catccggtga aagcactgga
     1201 ttttcgcacc cccttgaagg ccaagattat caatatctgt atttggctcc tgtcctcatc
     1261 cgtcggaatt tccgccattg tcttgggtgg tacaaaggtg agggaggacg tggacgttat
     1321 agagtgcagc ctgcagttcc ctgacgatga ctacagctgg tgggaccttt ttatgaagat
     1381 ctgtgtcttc atttttgctt tcgtcatccc cgtgctgatt atcatagtgt gctatacact
     1441 gatgatattg cggctgaaat ctgtgcgcct gctgagcggc agccgcgaaa aggatcggaa
     1501 tctgcgcttc atcacacgcc tcgtgctggt tgtagtcgct gtcttcgtgg tatgctggac
     1561 cccaatccac attttcatcc tggtggaagc tctgggatcc acctctcact ccactgccgc
     1621 acttagctca tattacttct gcattgcctt gggctacact aattcctcac tgaatcccat
     1681 tctgtacgca tttcttgacg agaacttcaa acggtgcttt agagactttt gcttccccct
     1741 caagatgcgc atggagcggc agtcaaccag ccgagtccgg aatacggtcc aggatcccgc
     1801 ctatctgcgg gacatcgacg gcatgaacaa gcctgtgcag ggcgcgccca tggtcgactc
     1861 atcacgtcgt aagtggatta aggcaggtca cgcagtcaga gctataggtc ggctgagctc
     1921 acccgtggtt tccgagcgga tgtaccccga ggacggcgtc ctgaagagcg agatcaagaa
     1981 ggggctgagg ctgaaggacg gcggccacta cgccgccgag gtcaagacca cctacaaggc
     2041 caagaagccc gtgcagctgc ccggcgccta catcgtcgac ataaagttgg acatcgtgtc
     2101 ccacaacgag gactacacca tcgtggaaca gtgcgaacgc gccgagggcc gccaccccac
     2161 cggcggcagg gacgagctgt acaagggagg tacaggaggg agtctggtga gcaagggcga
     2221 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     2281 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgaggcctt
     2341 tcagaccgct aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2401 gtcccctcag ttcacatacg gctccaaggc ctacattaag cacccagccg acatccccga
     2461 ctactttaag ctgtccttcc ccgagggctt caggtgggag cgcgtgatga acttcgagga
     2521 cggcggcatt attcacgtta accaggactc ctccctgcag gacggcgtat tcatctacaa
     2581 ggtgaagctg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat
     2641 gggctgggag gctacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc
     2701 tttctcccta tttgacaagg acggggatgg gacgataaca accaaggagc tggggacggt
     2761 gttgcggtct ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt
     2821 agatgccgac ggtgacggca cattcgactt ccctgagttc ctgacgatga tggcaagaag
     2881 aatgaatgac acagacagtg aagtggaaat tagagaagcg ttccgcgtgt ttgataatga
     2941 cggcaatggc tacatcggcg cagcagagct tcgccacgtg atgacagacc ttggagagaa
     3001 gttaacagat gaggaggttg atgaaatgat cagggtagca gacatcgatg gggatggtca
     3061 ggtaaactac gaagagtttg tacaaatgat gacagcgaag gatatcatgg ctcaagtcca
     3121 acttgtagag tctggaggag gtctcgtgcg gcccggcggt agcctgcgac tctcctgtgt
     3181 tgattcagag aggacgagct accctatggg gtggtttaga cgggcacccg gaaaggagcg
     3241 cgaattcgtt gcaagtatca cgtggagtgg cattgacccg acttatgctg atagcgttgc
     3301 cgaccggttt actacgtcca gagacgttgc gaataacacg ctgtacctcc agatgaactc
     3361 tctcaagcac gaagatacag cggtctatta ctgtgctgca agagcacccg tcggtcaatc
     3421 ttcctctcct tatgattatg actattgggg acaggggact caggtaactg tgtccagtta
     3481 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3541 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3601 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3661 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3721 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3781 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3841 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3901 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3961 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4021 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4081 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     4141 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4201 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4261 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     4321 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     4381 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     4441 ctctgcgcgc tcgctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4561 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5041 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5281 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5341 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5401 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5461 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5521 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5581 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5641 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5701 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5761 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5821 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5881 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5941 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6001 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6061 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6121 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6181 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6241 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6361 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     6421 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6481 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6541 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6601 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6661 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6721 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6781 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6841 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6901 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     6961 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     7021 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
     7081 taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7141 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     7201 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     7261 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     7321 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     7381 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     7441 tgggggcggg gggggggggg gggggggcgc cnnggggggg gggggggggg gggggggggg
     7501 gggggggggg gggggggggg gggggggggg cnnncccrmm mgmggggssc ksncscgynn
     7561 wybtntttyt ttttttgsgg gkgkggsggb cgbcgrrsmy sgayawacwc ggaacgcgcg
     7621 gggggggggg ggcggtggcg cggcgctctc ccccttatta gttcctgtgc gttctgtgtg
     7681 gttgggtgcg ttgcgggggc tggaggagag cgggtggggg gcggggatgc ccggggtggg
     7741 gggggggggg gggggggggc gggacggggg ggggggttgc ggctgggggg gggccggggc
     7801 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     7861 catgccttct tctttttcct acagctcctg ggcaacgtgc tggtt
//
