LOCUS       sequence-433961-        5505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KD Recombinase dependent conditional smGdP-10XOllas cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5505)
  AUTHORS   Sanfilippo P, Kim AJ, Bhukel A, Yoo J, Mirshahidi PS, Pandey V, 
            Bevir H, Yuen A, Mirshahidi PS, Guo P, Li HS, Wohlschlegel JA, Aso 
            Y, Zipursky SL
  TITLE     Mapping of multiple neurotransmitter receptor subtypes and distinct 
            protein complexes to the connectome.
  JOURNAL   Neuron. 2024 Jan 13:S0896-6273(23)00973-X. doi: 
            10.1016/j.neuron.2023.12.014.
  PUBMED    38262414
REFERENCE   2  (bases 1 to 5505)
  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: "Neuron. 
            2024 Jan 13:S0896-6273(23)00973-X. doi: 
            10.1016/j.neuron.2023.12.014."
COMMENT     Sequence Label: 
            pBS-KDRT-STOP-loxP-3XP3::dsRed-loxP-STOP-KDRT-smGdP-10XOllas
FEATURES             Location/Qualifiers
     source          1..5505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1090..1123
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1349..1358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1355..2032
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1485..1503)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1535..1555)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1753..1772
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1947..1970
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    2139..2172
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2187..2321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2224..2243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2278..2297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             2433..2474
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2484..2525
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2535..2576
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3108..3149
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3159..3200
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3219..3260
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3270..3311
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3525..3566
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3576..3617
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3627..3668
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     primer_bind     complement(3712..3731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3713..3731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3741..3758)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3741..3757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3750..3772)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3898..4353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4247..4266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4485..4553,4554..5345)
                     /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     complement(4703..4722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 ggaaacgata tcagacattt gtctgataat gcttcattat cagacaaatg tctgatatcg
      721 ttttaacgta agctagctag accggtgtcg actaaagcca aatagaaaat tattcagttc
      781 ctggcttaag tttttaaaag tgatattatt tatttggttg taaccaacca aaagaatgta
      841 aataactaat acataattat gttagtttta agttagcaac aaattgattt tagctatatt
      901 agctacttgg ttaataaata gaatatattt atttaaagat aattgcgttt ttattgtcag
      961 ggagtgagtt tgcttaaaaa ctcgtttagg tttgtcctcc cgaaattatt tatttaaatg
     1021 cgatggagag ttggcgccga atcgaaaact ttacgcgctt aaaagcacga gttggcatcc
     1081 ctaacgcgta taacttcgta taatgtatgc tatacgaagt tatcgtacgg gatctaattc
     1141 aattagagac taattcaatt agagctaatt caattaggat ccaagcttat cgatttcgaa
     1201 ccctcgaccg ccggagtata aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa
     1261 acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc gcagctgaac
     1321 aagctaaaca atcggctcga agccggtcgc caccatggcc tcctccgagg acgtcatcaa
     1381 ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat
     1441 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1501 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttcc agtacggctc
     1561 caaggtgtac gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga
     1621 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     1681 ggactcctcc ctccaggacg gctccttcat ctacaaggtg aagttcatcg gcgtgaactt
     1741 cccctccgac ggccccgtaa tgcagaagaa gactatgggc tgggaggcgt ccaccgagcg
     1801 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac aaggccctga agctgaagga
     1861 cggcggccac tacctggtgg agttcaagtc catctacatg gccaagaagc ccgtgcagct
     1921 gcccggctac tactacgtgg actccaagct ggacatcacc tcccacaacg aggactacac
     1981 catcgtggag cagtacgagc gcgccgaggg ccgccaccac ctgttcctgt aggggccgcg
     2041 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2101 cccacacctc cccctgaacc tgaaacataa aatgaatgat aacttcgtat aatgtatgct
     2161 atacgaagtt atcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2221 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat cactagtgat ctggccggaa
     2341 acgatatcag acatttgtct gataatgctt cattatcaga caaatgtctg atatcgtttg
     2401 gtggcggcgg ttcgggagga ggtggatcca tgagcggatt tgctaatgaa ctgggtccac
     2461 gacttatggg aaaacagcaa cagtcgggct tcgccaatga attgggcccg aggcttatgg
     2521 gcaaacaaca acagtccgga ttcgcaaacg agctcggacc tcgtttgatg ggcaagggtc
     2581 aaggcgtgtc gaagggcgag gagctgttta ctggtgtcgt tccaattttg gtggagttgg
     2641 atggcgatgt caatggacac aagtttagcg tacgtggaga gggtgaaggc gacgccacga
     2701 atggtaagct gactcttaag ttcatatgca ctacgggtaa gctgccggtg ccatggccca
     2761 cgctcgtcac gacgctggga ggaggcgtac aatgtttctc ccgataccct gaccatatga
     2821 agcagcacga cttcttcaaa agtgccatgc cagaaggata cgttcaggag cgcaccattt
     2881 cgtttaagga tgatggaact tacaagacac gagcagaggt gaaatttgag ggtgatacac
     2941 tggtaaatcg catagaactg aagggaattg atttcaagga ggatggcaac atcctgggcc
     3001 acaagttgga gtataatttc aacagccata atgtttacat aactgcggac aagcagaaaa
     3061 acggtataaa agcaaatttc aagatccgac acaatgtgga aggcggtagc ggcttcgcca
     3121 acgagctggg accgcgactg atgggcaagc aaggtcagag cggtttcgcc aacgagttgg
     3181 gaccccgcct gatgggtaag ggcggtcagc agggcggttc cggcttcgcc aatgagctcg
     3241 gcccaaggct tatgggaaaa caaggacaga gtggatttgc aaacgaactt ggtccgcgct
     3301 tgatgggaaa aggtggtgat ggtagcgtgc aacttgctga ccattaccag cagaataccc
     3361 caatcggaga cggacctgtg cttctgccag acaatcatta cttgagcacc caaagtgtgt
     3421 tgtccaaaga ccccaacgag aaacgcgatc acatggtgtt gctggagttc gtgaccgctg
     3481 cgggtattac tctgggtatg gatgagctct acaagggcca gggctccgga ttcgcgaacg
     3541 agctgggacc gaggctgatg ggcaagcaac agcagtccgg atttgccaac gaactgggac
     3601 cccggctcat gggcaagcag cagcaaagcg gcttcgcaaa tgagctcgga ccacgtctga
     3661 tgggaaagac atgttctaga gcggccgcca ccgcggtgga gctccaattc gccctatagt
     3721 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3781 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3841 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa
     3901 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4321 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4501 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4561 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4621 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4681 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4741 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4801 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4861 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4921 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4981 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5041 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5101 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5161 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5221 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5281 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5341 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5401 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatca
//
