LOCUS       sequence-377934-        7363 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 7363)
  AUTHORS   Kroning K, Wang W
  TITLE     Wang Lab opioid sensor
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7363)
  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            "unpublished"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..7363
                     /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             2349..2405
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWIKAGHAVRAIGRLSS"
     primer_bind     3101..3120
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3978..4566
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4031..4051)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4449..4460)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4609..4730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4620..4639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4674..4693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5109..5697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5189..5208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5868..6659,6660..6728))
                     /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     6491..6510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6729..6833)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6859..7314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6996..7017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7208..7227
                     /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 tcattttggg
     1141 caaagaattg gatccccacc atgagacccc agatcctgct gctcctggcc ctgctgaccc
     1201 taggcctggc tgactacaag gacgatgatg acgccatgga ctcttccgct gctcccacca
     1261 atgcctcaaa ttgtacagac gccttggcct atagctcttg ctctcccgca ccatctccgg
     1321 gaagctgggt taacctcagt catctcgacg ggaacctgtc cgacccttgt ggccctaata
     1381 ggaccgacct cggtggtcga gatagcctgt gcccccctac tggaagcccc agcatgatca
     1441 cagctatcac aataatggct ctctatagta tcgtctgcgt cgtgggactc tttggcaatt
     1501 ttctcgtcat gtacgttatc gtcagataca caaagatgaa aaccgcaacc aatatttaca
     1561 tctttaatct ggccctggcg gatgcccttg caaccagtac actgccgttt caatccgtga
     1621 attatctgat gggcacatgg cccttcggaa ccattctgtg caaaatcgta atctcaattg
     1681 attattataa catgttcacc tccattttta ccttgtgtac aatgtcagta gaccgctata
     1741 tcgccgtatg ccaccctgtc aaagctcttg atttcaggac ccctaggaat gccaagatca
     1801 tcaatgtgtg caattggatt ctgtctagcg ctatcggctt gccggtgatg ttcatggcca
     1861 ctaccaagta cagacaaggt tcaatcgact gtaccctcac attttcacat cccacgtggt
     1921 attgggagaa cttgctgaag atctgtgtgt tcatcttcgc cttcatcatg ccagtactga
     1981 taatcaccgt ttgttatggt cttatgatcc tgcgcctgaa gagcgtgcgc atgctcagcg
     2041 gctccaaaga aaaggaccgg aaccttcggc gaatcacacg gatggtcctg gtggttgtgg
     2101 ctgtgttcat tgtttgctgg acccctattc atatttatgt tataattaag gccctggtga
     2161 ccatcccgga aaccacattc cagaccgttt cctggcactt ttgtatagca ttgggatata
     2221 ccaacagttg ccttaatccc gttctgtacg cgtttctgga cgagaatttc aagcgatgct
     2281 tcagggagtt ctgcttcccc ctcaagatgc gcatggagcg gcagggcgcg cccatggtcg
     2341 actcatcacg tcgtaagtgg attaaggcag gtcacgcagt cagagctata ggtcggctga
     2401 gctcacccgt ggtttccgag cggatgtacc ccgaggacgg cgtcctgaag agcgagatca
     2461 agaaggggct gaggctgaag gacggcggcc actacgccgc cgaggtcaag accacctaca
     2521 aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt cgacataaag ttggacatcg
     2581 tgtcccacaa cgaggactac accatcgtgg aacagtgcga acgcgccgag ggccgccacc
     2641 ccaccggcgg cagggacgag ctgtacaagg gaggtacagg agggagtctg gtgagcaagg
     2701 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     2761 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2821 cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2881 tcctgtcccc tcagttcaca tacggctcca aggcctacat taagcaccca gccgacatcc
     2941 ccgactactt taagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg
     3001 aggacggcgg cattattcac gttaaccagg actcctccct gcaggacggc gtattcatct
     3061 acaaggtgaa gctgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
     3121 ccatgggctg ggaggctacg cgtgaccaac tgactgaaga gcagatcgca gaatttaaag
     3181 aggctttctc cctatttgac aaggacgggg atgggacgat aacaaccaag gagctgggga
     3241 cggtgttgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg
     3301 aagtagatgc cgacggtgac ggcacattcg acttccctga gttcctgacg atgatggcaa
     3361 gaagaatgaa tgacacagac agtgaagtgg aaattagaga agcgttccgc gtgtttgata
     3421 atgacggcaa tggctacatc ggcgcagcag agcttcgcca cgtgatgaca gaccttggag
     3481 agaagttaac agatgaggag gttgatgaaa tgatcagggt agcagacatc gatggggatg
     3541 gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaaggatatc atggctcaag
     3601 tccaacttgt agagtctgga ggaggtctcg tgcggcccgg cggtagcctg cgactctcct
     3661 gtgttgattc agagaggacg agctacccta tggggtggtt tagacgggca cccggaaagg
     3721 agcgcgaatt cgttgcaagt atcacgtgga gtggcattga cccgacttat gctgatagcg
     3781 ttgccgaccg gtttactacg tccagagacg ttgcgaataa cacgctgtac ctccagatga
     3841 actctctcaa gcacgaagat acagcggtct attactgtgc tgcaagagca cccgtcggtc
     3901 aatcttcctc tccttatgat tatgactatt ggggacaggg gactcaggta actgtgtcca
     3961 gttagaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4021 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4081 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4141 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4201 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4261 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4321 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4381 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4441 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4501 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4561 ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg
     4621 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4681 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4741 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     4801 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     4861 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4921 ccctctctgc gcgctcgctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5161 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5221 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5281 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5341 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5401 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5461 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5521 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5581 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5641 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5701 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5761 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5821 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5881 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5941 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6001 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6061 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6121 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6181 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6241 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6301 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6361 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6421 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6481 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6541 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6601 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6661 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6721 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6781 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6841 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6901 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6961 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     7021 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7081 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7141 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7201 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7261 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     7321 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcg
//
