LOCUS       sequence-418197-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent expression of mCherry tagged KASH protein domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  TITLE     Zweifel lab KASH plasmids
REFERENCE   2  (bases 1 to 4968)
  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"
COMMENT     Sequence Label: pAAV-FLEXfrt-mCherry-KASH
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    638..671
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1013..1720)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1288..1307)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1557..1575
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1861..1878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1867..2074
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2089..2229
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2089..2218
                     /label=AAV2 ITR
     rep_origin      2304..2759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2391..2410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2601..2622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2776..2795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2895..2917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2955..2973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3041..3145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3146..3214,3215..4006)
                     /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(3364..3383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4177..4765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4666..4685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4827..4956
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcgaggcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      601 gctaactacc ggtgctagcg tcgacagaat tggatccgaa gttcctattc tctagaaagt
      661 ataggaactt ccaaggcctg aagttcctat tcttcaaaag gtataggaac ttcgcggccg
      721 cttgatatct tggcgcgccc taggtgggag gtggcccgtt ggtgtaccgc agcatggggt
      781 agaaggatcg ggcaaagttg ttggcctggg tgcagctgta gtcatcttca gaggcaggta
      841 gcaggcaggc caggagcagc agcagcagca gaagcagctg caacggtagc gctgccctga
      901 tcacccttga gaggaaggag cgtctcggct gggagctgcc ggggctgtcg aggtggggca
      961 tcctgctgtc tgtctcctcc tcctcctctc gagatctgag tccggactta agcttgtaca
     1021 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     1081 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     1141 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     1201 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     1261 gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     1321 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     1381 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     1441 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     1501 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     1561 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     1621 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     1681 tgatgatggc catgttatcc tcctcgccct tgctcaccat caattggaag ttcctattct
     1741 ttctagagta taggaacttc ctgcagtagg ctagcgaagt tcctattcct tttgaagtat
     1801 aggaacttcg tcgagggggg gcccgagctt atcgatgaat tcctagagct cgctgatcag
     1861 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1921 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1981 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2041 aggattggga agagaatagc aggcatgctg gggaggtacc tcggccgcag gaacccctag
     2101 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2161 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2221 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2281 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2641 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2701 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     2761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     2821 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2881 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2941 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3001 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3061 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3121 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3181 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3241 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3301 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3361 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3421 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3481 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3541 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3601 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3661 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3721 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3781 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3841 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     3901 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3961 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4021 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4081 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4141 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4201 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4261 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4321 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4381 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4441 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4501 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4561 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4621 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4681 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4741 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4801 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
     4861 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     4921 gagagggagt ggccaactcc atcactaggg gttcctgcgg cctctaga
//
