LOCUS       sequence-367150-        8144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible disruption of LINC Complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  TITLE     Lammerding Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8144)
  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: BFP-KASH2
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    224..305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(261..280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     315..334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        434..1612
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          665..1603
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1560..1580
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(1670..2287,2288..2293,2294..2329,2330..2332,2333..2368,
                     2369..2371,2372..2407,2408..2416)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(2435..3009,3010..3012,3013..3021)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2602..2619)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2829..2848
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3022..3825
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3085..3104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3695..3714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3833..3850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3839..4063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4252..4314
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   6147..6181
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    6491..6509
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6533..6551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6575..6593
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6617..6635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6659..6677
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6701..6719
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6752..6790
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6787..6811
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          7022..7154
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7137..7156
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             7219..7914
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
ORIGIN
        1 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
       61 atcaatcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
      121 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
      181 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
      241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      301 actgcattct agttgtggtt tgtccaaact catcaatgct cgacgataag ctttgcaaag
      361 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
      421 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
      481 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt
      541 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      601 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      661 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
      721 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
      781 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
      841 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
      901 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
      961 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     1021 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     1081 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     1141 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     1201 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     1261 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     1321 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     1381 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     1441 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     1501 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     1561 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaatttc
     1621 gacatttaaa tttaattaat ctcgacggta tcggttggga tctgccacca tgtccagact
     1681 ggacaagagc aaagtcataa acggcgctct ggaattactc aatggagtcg gtatcgaagg
     1741 cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca
     1801 cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca
     1861 tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa
     1921 gtcattccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg
     1981 cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg
     2041 cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg
     2101 cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga
     2161 ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccggcagg gagccgaacc
     2221 tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga
     2281 aagcggcggg ccggccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc
     2341 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga
     2401 catgctcccc ggctaaccca aacccgctga tccgcccctc tccctccccc ccccctaacg
     2461 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     2521 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2581 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2641 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2701 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2761 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2821 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2881 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2941 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3001 gatgataata tggccacaac catgggatcg gccattgaac aagatggatt gcacgcaggt
     3061 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3121 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3181 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3241 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3301 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3361 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3421 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3481 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3541 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3601 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3661 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3721 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3781 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgatgatc agcctcgact
     3841 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3901 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3961 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4021 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcatcgat gctagttaaa
     4081 agttttgtta ctttatagaa gaaattttga gtttttgttt ttttttaata aataaataaa
     4141 cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt
     4201 aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca
     4261 attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct agggttaatc
     4321 tagtatacgc gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6121 cccgaaaagt gccaccgcta gattaaccct agaaagatag tctgcgtaaa attgacgcat
     6181 gcattcttga aatattgctc tctctttcta aatagcgcga atccgtcgct gtgcatttag
     6241 gacatctcag tcgccgcttg gagctcccgt gaggcgtgct tgtcaatgcg gtaagtgtca
     6301 ctgattttga actataacga ccgcgtgagt caaaatgacg catgattatc ttttacgtga
     6361 cttttaagat ttaactcata cgataattat attgttattt catgttctac ttacgtgata
     6421 acttattata tatatatttt cttgttatag atatcgggtc agcttggcct agcgctgtcg
     6481 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
     6541 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
     6601 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     6661 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     6721 aagtgaaagt cgagctcggt acccgggtcg aggtaggcgt gtacggtggg aggcctatat
     6781 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     6841 cctccataga agacaccggg accgatccag cctccgcgga atttgattag ctttattgcg
     6901 gtagtttatc acagttaaat tgctaacgca gtcagtgctt ctgacacaac agtctcgaac
     6961 ttaagctgca gtgactctct taaggtagcc ttgcagaagt tggtcgtgag gcactgggca
     7021 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     7081 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     7141 ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaaggct agaggatccc
     7201 accatggcta gcggcggaag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
     7261 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
     7321 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     7381 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
     7441 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
     7501 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     7561 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     7621 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     7681 ggcagaaacg acatggccct gaagcttgtg ggcgggagcc atctgatcgc aaacgccaag
     7741 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     7801 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     7861 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taatggcgga
     7921 cctaggtccg gactcagatc tagctcccag cctagacgat ccttcctctc aagggtgatc
     7981 agggcagcgc taccgttgca gctgcttctg ctgctgctgc tgctcctggc ctgcctgcta
     8041 cctgcctctg aagatgacta cagctgcacc caggccaaca actttgcccg atccttctac
     8101 cccatgctgc ggtacaccaa cgggccacct cccacctagg aatt
//
