LOCUS       sequence-377865-        7562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent-based opioid sensor for the kappa 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 7562)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7562)
  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            "unpublished"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
FEATURES             Location/Qualifiers
     source          1..7562
                     /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             2399..2455
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWIKAGHAVRAIGRLSS"
     primer_bind     3151..3170
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4028..4616
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4081..4101)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4499..4510)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4659..4780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4670..4689)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4724..4743
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4942..5071
                     /label=AAV2 ITR
     rep_origin      complement(5308..5896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5388..5407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6067..6858,6859..6927))
                     /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     6690..6709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6928..7032)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7058..7513)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7195..7216)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7407..7426
                     /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 ccgaatcaca 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 cccatggtcg actcatcacg
     2401 tcgtaagtgg attaaggcag gtcacgcagt cagagctata ggtcggctga gctcacccgt
     2461 ggtttccgag cggatgtacc ccgaggacgg cgtcctgaag agcgagatca agaaggggct
     2521 gaggctgaag gacggcggcc actacgccgc cgaggtcaag accacctaca aggccaagaa
     2581 gcccgtgcag ctgcccggcg cctacatcgt cgacataaag ttggacatcg tgtcccacaa
     2641 cgaggactac accatcgtgg aacagtgcga acgcgccgag ggccgccacc ccaccggcgg
     2701 cagggacgag ctgtacaagg gaggtacagg agggagtctg gtgagcaagg gcgaggagga
     2761 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2821 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg cctttcagac
     2881 cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2941 tcagttcaca tacggctcca aggcctacat taagcaccca gccgacatcc ccgactactt
     3001 taagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg
     3061 cattattcac gttaaccagg actcctccct gcaggacggc gtattcatct acaaggtgaa
     3121 gctgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg
     3181 ggaggctacg cgtgaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc
     3241 cctatttgac aaggacgggg atgggacgat aacaaccaag gagctgggga cggtgttgcg
     3301 gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc
     3361 cgacggtgac ggcacattcg acttccctga gttcctgacg atgatggcaa gaagaatgaa
     3421 tgacacagac agtgaagtgg aaattagaga agcgttccgc gtgtttgata atgacggcaa
     3481 tggctacatc ggcgcagcag agcttcgcca cgtgatgaca gaccttggag agaagttaac
     3541 agatgaggag gttgatgaaa tgatcagggt agcagacatc gatggggatg gtcaggtaaa
     3601 ctacgaagag tttgtacaaa tgatgacagc gaaggatatc atggctcaag tccaacttgt
     3661 agagtctgga ggaggtctcg tgcggcccgg cggtagcctg cgactctcct gtgttgattc
     3721 agagaggacg agctacccta tggggtggtt tagacgggca cccggaaagg agcgcgaatt
     3781 cgttgcaagt atcacgtgga gtggcattga cccgacttat gctgatagcg ttgccgaccg
     3841 gtttactacg tccagagacg ttgcgaataa cacgctgtac ctccagatga actctctcaa
     3901 gcacgaagat acagcggtct attactgtgc tgcaagagca cccgtcggtc aatcttcctc
     3961 tccttatgat tatgactatt ggggacaggg gactcaggta actgtgtcca gttagaagct
     4021 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4081 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4141 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4201 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4261 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4321 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4381 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4441 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4501 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4561 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4621 ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4681 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4741 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4801 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     4861 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     4921 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4981 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     5041 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     5101 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5161 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6001 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6061 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6121 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6181 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6241 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6301 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6361 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6421 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6481 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6541 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6601 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6661 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6721 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6781 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6841 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6901 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7021 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     7081 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7141 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7201 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7261 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7321 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7381 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7441 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7501 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7561 cg
//
