LOCUS       sequence-377952-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green 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 7043)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7043)
  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            "unpublished"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-SPOTIT(R271F)
FEATURES             Location/Qualifiers
     source          1..7043
                     /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     2590..2611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2717..2738)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2978..2997)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3509..4097
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3562..3582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3980..3991)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4140..4261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4151..4170)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4205..4224
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4423..4552
                     /label=AAV2 ITR
     rep_origin      complement(4789..5377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4869..4888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5548..6339,6340..6408))
                     /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     6171..6190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6409..6513)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6539..6994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6676..6697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6888..6907
                     /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 tgaagacgat catcgccctg agctacatct tctgcctggt
     1201 gttcgccgac tacaaggacg atgatgacgc catggacagc cccattcaaa tcttccgggg
     1261 cgagcccgga ccaacctgcg ctccaagcgc ttgcctgcca ccgaattcta gtgcatggtt
     1321 cccagggtgg gcagagcccg atagcaatgg ctcagccggg tccgaggatg cgcagcttga
     1381 acccgctcac atctcacccg ctatccctgt cattattacg gcagtctact cagttgtttt
     1441 cgttgtgggc ctggttggaa atagcctcgt gatgtttgtg attatcaggt atactaaaat
     1501 gaagaccgcc acgaatatct acatctttaa cctggctctg gctgacgcct tggtgaccac
     1561 cactatgcca tttcagtcca ccgtctatct tatgaactct tggccattcg gcgacgttct
     1621 gtgcaagatt gtcatttcca ttgactatta taatatgttt acatcaatct ttacgcttac
     1681 aatgatgagc gtggaccgat acattgctgt ttgccatccg gtgaaagcac tggattttcg
     1741 cacccccttg aaggccaaga ttatcaatat ctgtatttgg ctcctgtcct catccgtcgg
     1801 aatttccgcc attgtcttgg gtggtacaaa ggtgagggag gacgtggacg ttatagagtg
     1861 cagcctgcag ttccctgacg atgactacag ctggtgggac ctttttatga agatctgtgt
     1921 cttcattttt gctttcgtca tccccgtgct gattatcata gtgtgctata cactgatgat
     1981 attgcggctg aaatctgtgc gcctgctgag cggcagccgc gaaaaggatc ggaatctgcg
     2041 cttcatcaca cgcctcgtgc tggttgtagt cgctgtcttc gtggtatgct ggaccccaat
     2101 ccacattttc atcctggtgg aagctctggg atccacctct cactccactg ccgcacttag
     2161 ctcatattac ttctgcattg ccttgggcta cactaattcc tcactgaatc ccattctgta
     2221 cgcatttctt gacgagaact tcaaacggtg ctttagagac ttttgcttcc ccctcaagat
     2281 gcgcatggag cggcagtcaa ccagccgagt ccggaatacg gtccaggatc ccgcctatct
     2341 gcgggacatc gacggcatga acaagcctgt gcagggcgcg cccaacgtct atatcaaggc
     2401 cgacaagcag aagaacggca tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg
     2461 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa
     2581 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2641 cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac
     2701 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2761 gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2821 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2881 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2941 cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3001 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3061 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctta aggaagatat
     3121 catggctcaa gtccaacttg tagagtctgg aggaggtctc gtgcggcccg gcggtagcct
     3181 gcgactctcc tgtgttgatt cagagaggac gagctaccct atggggtggt ttagacgggc
     3241 acccggaaag gagcgcgaat tcgttgcaag tatcacgtgg agtggcattg acccgactta
     3301 tgctgatagc gttgccgacc ggtttactac gtccagagac gttgcgaata acacgctgta
     3361 cctccagatg aactctctca agcacgaaga tacagcggtc tattactgtg ctgcaagagc
     3421 acccgtcggt caatcttcct ctccttatga ttatgactat tggggacagg ggactcaggt
     3481 aactgtgtcc agttagaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3541 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3601 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3661 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3721 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3781 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3841 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3901 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3961 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4021 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4081 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
     4141 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4261 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4321 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     4381 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4441 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4501 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta
     4561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4981 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5221 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5341 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5401 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5461 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5521 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5581 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5641 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5701 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5761 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5821 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5881 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5941 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6001 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6061 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6121 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6181 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6241 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6301 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6361 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6421 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6481 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     6541 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6601 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     6661 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6721 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     6781 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6841 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6901 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6961 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     7021 tgttgggaag ggcgatcggt gcg
//
