LOCUS       sequence-122540-        8010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nesprin headless control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8010)
  AUTHORS   Arsenovic PT, Ramachandran I, Bathula K, Zhu R, Narang JD, Noll NA, 
            Lemmon CA, Gundersen GG, Conway DE
  TITLE     Nesprin-2G, a Component of the Nuclear LINC Complex, Is Subject to 
            Myosin-Dependent Tension.
  JOURNAL   Biophys J. 2016 Jan 5;110(1):34-43. doi: 10.1016/j.bpj.2015.11.014.
  PUBMED    26745407
REFERENCE   2  (bases 1 to 8010)
  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.bpj.2015.11"; journalName: "Biophys J"; date: "2016-01-5-
            5"; volume: "110"; issue: "1"; pages: "34-43"
COMMENT     Sequence Label: pcdna nesprin HL
FEATURES             Location/Qualifiers
     source          1..8010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             927..1634
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(1755..1757,1758..1760,1761..2471)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1800..1821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2061..2080)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2408..2429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3604..3621)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3610..3834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3880..4308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4322..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4564..4583
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4718..5512
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4772..4791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5382..5401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5686..5807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5723..5742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5777..5796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5856..5872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5856..5872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5869..5891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5880..5896
                     /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)."
     promoter        complement(join(5904..5910,5911..5928,5929..5934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5949..5970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6087..6104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6258..6843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6338..6357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7014..7805,7806..7874))
                     /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     7637..7656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7875..7979)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagatgg tgagcaaggg cgaggagacc acaatgggcg
      961 taatcaagcc cgacatgaag atcaagctga agatggaggg caacgtgaat ggccacgcct
     1021 tcgtgatcga gggcgagggc gagggcaagc cctacgacgg caccaacacc atcaacctgg
     1081 aggtgaagga gggagccccc ctgcccttct cctacgacat tctgaccacc gcgttcgcct
     1141 acggcaacag ggccttcacc aagtaccccg acgacatccc caactacttc aagcagtcct
     1201 tccccgaggg ctactcttgg gagcgcacca tgaccttcga ggacaagggc atcgtgaagg
     1261 tgaagtccga catctccatg gaggaggact ccttcatcta cgagatacac ctcaagggcg
     1321 agaacttccc ccccaacggc cccgtgatgc agaagaagac caccggctgg gacgcctcca
     1381 ccgagaggat gtacgtgcgc gacggcgtgc tgaagggcga cgtcaagcac aagctgctgc
     1441 tggagggcgg cggccaccac cgcgttgact tcaagaccat ctacagggcc aagaaggcgg
     1501 tgaagctgcc cgactatcac tttgtggacc accgcatcga gatcctgaac cacgacaagg
     1561 actacaacaa ggtgaccgtt tacgagagcg ccgtggcccg caactccacc gacggcatgg
     1621 acgagctgta caaggggcca ggtggtgcag ggccaggtgg tgcagggcca ggtggtgcag
     1681 ggccaggtgg tgcagggccc ggtggtgcag gtccaggtgg tgcaggtcca ggtggtgcag
     1741 gtccaggtgg tgctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1801 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1861 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     1921 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     1981 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2041 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2101 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2161 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     2221 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2281 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2341 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     2401 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2461 agctgtacaa ggcggccgcg caggagcttc acagcaagct caggctcaaa cagaccgtgc
     2521 agcagctcaa gtctgacatc ggcagcatcg ctgcttggct tgggaaaact gaggccgagc
     2581 tggaagctct gaagttggcc gagcctccct ctgacatcca ggaaatagcg ctcagggtga
     2641 agagactgca agagatacta aaagccttcg atacctacaa ggccttgatg gtctccgtca
     2701 atgtgagcca caaggaatat ctgccaagcc agagccccga agccacagaa ctccaaaata
     2761 gactccatca gctgagcctg tcctgggact cggtccaggg cgtattggac agctggcgag
     2821 gagacttacg gcagtcactc atgcagtgcc aggactttca ccagctgagt caagacttgc
     2881 tcctgtggct agcgacagca gagagccgca ggcagaaggc gcatgtcacc agcccagagg
     2941 ccgaccgcca ggtcctcctg gagtgtcaga aagacctaat gcgcctagaa aaggagttgg
     3001 tagcccgtca gcctcaagtc agctccttgc gggagatttc ctccagcctt ctggtgaagg
     3061 ggcaaggaga agactacatt gaggccgaag agaaggtcca cgtgatcgag aagaaactca
     3121 agcaactgca agagcaagtg gctcaagatc tgatgtcctt gcagagaagc ctggatccgg
     3181 atgcttctct aaccagcttt gatgaggtgg actcaggaga acagctccca gcagcctttg
     3241 caaagagtcc aaggccccgc tggaccttct tggaagagga agaggaggag gaggagacag
     3301 acagcaggat gccccacctc gacagccccg gcagctccca gccgagacgc tccttcctct
     3361 caagggtgat cagggcagcg ctaccgttgc agctgcttct gctgctgctg ctgctcctgg
     3421 cctgcctgct acctgcctct gaagatgact acagctgcac ccaggccaac aactttgccc
     3481 gatccttcta ccccatgctg cggtacacca acgggccacc tcccacctag accggtgaat
     3541 tccaccacac tggactagtg gatccgagct cggtaccaag cttaagttta aaccgctgat
     3601 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3661 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3721 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3781 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3841 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4681 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4741 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4801 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4861 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4921 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4981 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5041 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5101 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5161 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5221 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5281 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5341 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5401 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5461 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5521 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5581 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5641 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5701 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5761 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5821 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5881 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5941 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6001 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6061 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6121 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6181 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6241 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6421 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6661 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6781 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6841 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6901 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6961 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7021 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7081 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7141 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7201 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7261 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7321 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7381 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7441 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7501 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7561 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7621 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7681 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7741 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7801 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7861 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7921 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7981 acatttcccc gaaaagtgcc acctgacgtc
//
