LOCUS       sequence-416789-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane tethered mCherry shRNA targeting Ctnnd2 N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  AUTHORS   Tan CX, Bindu DS, Hardin EJ, Sakers K, Baumert R, Ramirez JJ, Savage
            JT, Eroglu C
  TITLE     delta-Catenin controls astrocyte morphogenesis via layer-specific 
            astrocyte-neuron cadherin interactions.
  JOURNAL   J Cell Biol. 2023 Nov 6;222(11):e202303138. doi: 
            10.1083/jcb.202303138. Epub 2023 Sep 14.
  PUBMED    37707499
REFERENCE   2  (bases 1 to 6992)
  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: "J Cell 
            Biol."; date: "2023-11-6"; volume: "222(11):e202303138. doi"; pages:
            " 10.1083/jcb.202303138. Epub 2023 Sep 14"
COMMENT     Sequence Label: pPB-shCtnnd2.1-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..814
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     566..586
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     737..756
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    911..1028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(1105..1138)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   1435..1497
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1535..1639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1640..1708,1709..2500)
                     /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(1858..1877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2671..3259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3160..3179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3413..3430
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3498..3632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3522..3541)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3576..3595
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(3638..3833)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3652..3787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    3892..3913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3928..3933,3934..3951,3952..3958)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3966..3982
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3971..3993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3990..4006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3990..4006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4029..4063
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     4716..4735
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4734..4789
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4835..4854
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(4958..5665)
                     /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(5233..5252)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5502..5520
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5748..5767)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5775..6791
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5829..5851)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(join(6792..6992,1..77))
                     /label=chicken beta-actin promoter
ORIGIN
        1 aataaataaa tacaaaattg ggggtgggga ggggggggag atggggagag tgaagcagaa
       61 cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt actgccaagt
      121 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
      181 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
      241 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
      301 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
      361 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
      421 aattgattac tattaataac tagtcaggcc cgaaggaata gaagaagaag gtggagagag
      481 agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta
      541 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
      601 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      661 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      721 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      781 ggctttatat atcttgtgga aaggacgaaa caccggccag gcttaaacac ctccaatctc
      841 gagattggag gtgtttaagc ctggtttttg aattctcgac ctcgagacaa atggcagtat
      901 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
      961 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     1021 aaaattttcg ggtttattac agggacagca gagatccact ttggcactag ttggactact
     1081 gcgccctaca gatcccttaa tcgcataact tcgtatagca tacattatac gaagttatgc
     1141 gattaataag tataatttgt ttctattatg tataagttaa gctaattact tattttataa
     1201 tacaacatga ctgtttttaa agtacaaaat aagtttattt ttgtaaaaga gagaatgttt
     1261 aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttta ataaataaat
     1321 aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa
     1381 cttaatatct attcaaatta ataaataaac ctcgatatac agatcgataa aacacatgcg
     1441 tcaattttac gcatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta
     1501 agtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1621 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1861 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2161 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2281 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2341 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2401 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2461 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3361 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3421 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3481 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
     3541 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3601 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
     3661 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     3721 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     3781 gcgggatggg cggagttagg ggcgggacta tggttgctga cttattgaga tgcggatccg
     3841 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     3901 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3961 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4021 gcttttaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt gaaatattgc
     4081 tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc agtcgccgct
     4141 tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt gaactataac
     4201 gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag atttaactca
     4261 tacgataatt atattgttat ttcatgttct acttacgtga taacttatta tatatatatt
     4321 ttcttgttat agatatcgtg actaatatat aataaaagca ttacacgtct tgagcgattg
     4381 tgtaggctgg agctctgcag gtcgagggat ctccataaga gaagagggac agctatgact
     4441 gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga aaattttagg
     4501 gatgttaaag aaaaaaataa cacaaaacaa aatataaaaa aaatctaacc tcaagtcaag
     4561 gcttttctat ggaataagga atggacagca gggggctgtt tcatatactg atgacctctt
     4621 tatagccaac ctttgttcat ggcagccagc atatgggcat atgttgccaa actctaaacc
     4681 aaatactcat tctgatgttt taaatgattt gccctcccat atgtccttcc gagtgagaga
     4741 cacaaaaaat tccaacacac tattgcaatg aaaataaatt tcctttatta gccagaagtc
     4801 agatgctcaa ggggcttcat gatgtcccca taatttttgg cagagggaaa aagatctcag
     4861 tggtatttgt gagccagggc attggccaca ccagccacca ccttctgata ggcagcctgc
     4921 acctgaggag tgcggccgct tacaggatga tgcagcactt gtacagctcg tccatgccgc
     4981 cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt
     5041 gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct
     5101 tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc
     5161 tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct
     5221 cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca
     5281 gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca
     5341 cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct
     5401 tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact
     5461 gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg
     5521 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
     5581 cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt
     5641 tatcctcctc gcccttgctc accataagct tgagctcgag atctgagtcc ggtagcgggc
     5701 ccgcggtacc gtcgactgca gaattctttg ccaaaatgat gagacagcac aataaccagc
     5761 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta
     5821 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag
     5881 ccgtcccccc gcggacagcc ccgaggctgg agagggagaa ggggacggcg gcgcggcgac
     5941 gcacgaaggc cctccccgcc catttccttc ctgccggcgc cgcaccgctt cgccccgcgc
     6001 ccgctagagg gggtgcggcg gcgcctccca gatttcggct ccgcacagat ttgggacaaa
     6061 ggaagtccct gcgccctctc gcacgattac cataaaaggc aatggctgcg gctcgccgcg
     6121 cctcgacagc cgccggcgct ccgggggccg ccgcgcccct cccccgagcc ctccccggcc
     6181 cgaggcggcc ccgccccgcc cggcaccccc acctgccgcc accccccgcc cggcacggcg
     6241 agccccgcgc cacgccccgt acggagcccc gcacccgaag ccgggccgtg ctcagcaact
     6301 cggggagggg ggtgcagggg gggttgcagc ccgaccgacg cgcccacacc ccctgctcac
     6361 ccccccacgc acacaccccg cacgcagcct ttgttcccct cgcagccccc cccgcaccgc
     6421 ggggcaccgc ccccggccgc gctcccctcg cgcacactgc ggagcgcaca aagccccgcg
     6481 ccgcgcccgc agcgctcaca gccgccgggc agcgcggagc cgcacgcggc gctccccacg
     6541 cacacacaca cgcacgcacc ccccgagccg ctccccccgc acaaagggcc ctcccggagc
     6601 ccctcaaggc tttcacgcag ccacagaaaa gaaacaagcc gtcattaaac caagcgctaa
     6661 ttacagcccg gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt
     6721 cagagccggg gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc
     6781 gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc
     6841 gccataaaag gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc
     6901 ctcgccccgc cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc cccccccccc
     6961 cccgccccca tcgctgcaca aaataattaa aa
//
