LOCUS       sequence-469976-        7438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Opto-MOR in a Cre dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7438)
  AUTHORS   Siuda ER, Copits BA, Schmidt MJ, Baird MA, Al-Hasani R, Planer WJ, 
            Funderburk SC, McCall JG, Gereau RW 4th, Bruchas MR
  TITLE     Spatiotemporal control of opioid signaling and behavior.
  JOURNAL   Neuron. 2015 May 20;86(4):923-35. doi: 10.1016/j.neuron.2015.03.066.
            Epub 2015 Apr 30.
  PUBMED    25937173
REFERENCE   2  (bases 1 to 7438)
  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"; doi: 
            "10.1016/j.neuron.2015.03.066"; journalName: "Neuron"; date: 
            "2015-05-20- 20"; volume: "86"; issue: "4"; pages: "923-35"
COMMENT     Sequence Label: pAAV-Ef1a-DIO-OMOR-eYFP
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     277..298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    352..385
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    436..469
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(500..1678)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          509..1447
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(532..552)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   1779..1908
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1970..2558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2050..2069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2729..3520,3521..3589))
                     /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     3352..3371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3590..3694)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3762..3780
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3818..3840)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3940..3959
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3976..4431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4113..4134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4325..4344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4506..4646
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4517..4646
                     /label=AAV2 ITR
     polyA_signal    4686..5162
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     4944..4963
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    5194..5782
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5300..5311
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5709..5729
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(5807..5840)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5891..5924)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      7056..7065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7062..7064,7065..7067,7068..7438,1..343)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7107..7128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7368..7387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
       61 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      121 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      181 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
      241 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      301 cgccgccggg atcactctcg gcatggacga gctgtacaag taaggcgcgc cataacttcg
      361 tataatgtat gctatacgaa gttattaaga ggtttcatat tgctaatagc agctacaatc
      421 cagctaccat tctgcataac ttcgtataaa gtatcctata cgaagttatt ccggagtcga
      481 ctctagagga tccggtacct cacgacacct gaaatggaag aaaaaaactt tgaaccactg
      541 tctgaggctt gagaatgaac caagatccaa actcaaaaag ggcaaattcc aaggagaatt
      601 acatcaagtg ccaagctggc ctaacttcag tctccaccca ctcagtgtgg ggaaactcca
      661 tcgcataaaa cccctccccc caacctaaag acgacgtact ccaaaagctc gagaactaat
      721 cgaggtgcct ggacggcgcc cggtactccg tggagtcaca tgaagcgacg gctgaggacg
      781 gaaaggccct tttcctttgt gtgggtgact cacccgcccg ctctcccgag cgccgcgtcc
      841 tccattttga gctccctgca gcagggccgg gaagcggcca tctttccgct cacgcaactg
      901 gtgccgaccg ggccagcctt gccgcccagg gcggggcgat acacggcggc gcgaggccag
      961 gcaccagagc aggccggcca gcttgagact acccccgtcc gattctcggt ggccgcgctc
     1021 gcaggccccg cctcgccgaa catgtgcgct gggacgcacg ggccccgtcg ccgcccgcgg
     1081 ccccaaaaac cgaaatacca gtgtgcagat cttggcccgc atttacaaga ctatcttgcc
     1141 agaaaaaaag cgtcgcagca ggtcatcaaa aattttaaat ggctagagac ttatcgaaag
     1201 cagcgagaca ggcgcgaagg tgccaccaga ttcgcacgcg gcggccccag cgcccaggcc
     1261 aggcctcaac tcaagcacga ggcgaagggg ctccttaagc gcaaggcctc gaactctccc
     1321 acccacttcc aacccgaagc tcgggatcaa gaatcacgta ctgcagccag gtggaagtaa
     1381 ttcaaggcac gcaagggcca taacccgtaa agaggccagg cccgcgggaa ccacacacgg
     1441 cacttacctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg
     1501 cacttatata cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac
     1561 tttcccagtt taccccgcgc caccttctct aggcaccggt tcaattgccg acccctcccc
     1621 ccaacttctc ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagccaa
     1681 ttcccactcc tttcaagacc tagaaggtcc attagctgca aagattcctc tctgtttaaa
     1741 actttatcca tctttgcaaa gcttacgcgt gcggccgcag gaacccctag tgatggagtt
     1801 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     1861 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
     1921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3601 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3661 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3721 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3781 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3841 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3901 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3961 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     4021 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
     4081 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4141 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4201 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4261 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4321 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
     4381 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
     4441 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4501 gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     4561 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     4621 ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtggt tacctacaaa
     4681 atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc
     4741 caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa
     4801 ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc
     4861 aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg
     4921 agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag
     4981 gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat aatattatag
     5041 aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc
     5101 actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac
     5161 ccgtagatct ctcgagcagc gctcggtatc gatgcgggga ggcggcccaa agggagatcc
     5221 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
     5281 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     5341 gaatccaggt ggcaacatag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca
     5401 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     5461 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     5521 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     5581 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     5641 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     5701 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     5761 ttttgtaatc cagaggttga ttatcgataa gcttgatatc gaattcataa cttcgtatag
     5821 catacattat acgaagttat tctttgcacc attctaaaga ataacagtga taatttctgg
     5881 gttaaggcaa ataacttcgt ataggatact ttatacgaag ttatgctagc atgaacggca
     5941 cagagggccc caatttttat gtgcccttct ccaacatcac gggcgtggtg cgcagcccct
     6001 ttgagcagcc gcagtactac ctggcggagc catggcagtt ctccatgctg gcagcctaca
     6061 tgttcctgct catcgtgctg ggcttcccca tcaacttcct cacgctctac gtcattgtaa
     6121 gatacaccaa aatgaagact ccactcaact acatcctgct caacttggct gtggctgacc
     6181 tcttcatggt cttcggagga ttcaccacca ccctctacac ctcactgcat ggctactttg
     6241 tctttgggcc cacaggctgc aaccttgagg gcttctttgc cacccttgga ggtgaaatcg
     6301 gcctgtggtc cctggtagtc ctggccattg agcgctacgt ggtggtctgc cacccagtca
     6361 aagccctgga tttccgtacc ccccgaaatg ccattatggg tgtggccttc acctgggtca
     6421 tggcgttggc ctgtgctgct cccccactgg ttggctggtc caggtacatc cccgagggca
     6481 tgcagtgttc atgtgggatt gactactata cactcaagcc tgaggtcaac aatgagtcct
     6541 tcgtcatcta catgttcgtg gtccacttca ccatccccat gatcgtcatc ttcttctgtt
     6601 acggcctgat gatcttacga ctcaagagcg ttcgcatgct atcgggctcc aaagaaaagg
     6661 acaggaatct gcgcaggatc acgcgcatgg tcatcatcat ggtcatcttc ttcctgatct
     6721 gctggcttcc ctatgccagt gtggccatgt acatctttac ccaccagggc tccaacttcg
     6781 gccccatctt catgaccctt cccgctttct ttgctaagac cgcctccatc tacaacccaa
     6841 tcatctacat catgatgaac aagcagttca gagagttctg catcccaacc tcgtccacga
     6901 tcgaacagca aaactccacg cgagtccgtc agaacactag ggaacatccc tccacggcta
     6961 atacagtgga tcgaactaac caccagctag aaaatctgga ggcagaaact gctccattgc
     7021 ccacggagac cagccaggtg gctccagccg ctagcgccac catggtgagc aagggcgagg
     7081 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     7141 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     7201 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     7261 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     7321 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     7381 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagct
//
