LOCUS       sequence-224387-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of lamin B1 with N-terminal Spot-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  TITLE     ChromoTek Spot-tag plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5739)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSpot-Tag-Lamin
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             562..597
                     /codon_start=1
                     /product="engineered Spot-Tag(R) peptide that binds with 
                     high affinity to a nanobody"
                     /label=Spot-Tag
                     /translation="PDRVRAVSHWSS"
     polyA_signal    2467..2588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2504..2523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2558..2577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2595..3050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2732..2753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2944..2963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3077..3181
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3183..3540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3183..3374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3206..3226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3391..3526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3575..4369
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3575..4369
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3629..3648)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4239..4258
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4557..4576)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4601..4648
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4977..5565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5466..5485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat gcctgatcgc gtgcgcgccg tgagccattg gagcagcgcg
      601 actgcgaccc ccgtgccgcc gcggatgggc agccgcgctg gcggccccac cacgccgctg
      661 agccccacgc gcctgtcgcg gctccaggag aaggaggagc tgcgcgagct caatgaccgg
      721 ctggcggtgt acatcgacaa ggtgcgcagc ctggagacgg agaacagcgc gctgcagctg
      781 caggtgacgg agcgcgagga ggtgcgcggc cgtgagctca ccggcctcaa ggcgctctac
      841 gagaccgagc tggccgacgc gcgacgcgcg ctcgacgaca cggcccgcga gcgcgccaag
      901 ctgcagatcg agctgggcaa gtgcaaggcg gaacacgacc agctgctcct caactatgct
      961 aagaaggaat ctgatcttaa tggcgcccag atcaagcttc gagaatatga agcagcactg
     1021 aattcgaaag atgcagctct tgctactgca cttggtgaca aaaaaagttt agagggagat
     1081 ttggaggatc tgaaggatca gattgcccag ttggaagcct ccttagctgc agccaaaaaa
     1141 cagttagcag atgaaacttt acttaaagta gatttggaga atcgttgtca gagccttact
     1201 gaggacttgg agtttcgcaa aagcatgtat gaagaggaga ttaacgagac cagaaggaag
     1261 catgaaacgc gcttggtaga ggtggattct gggcgtcaaa ttgagtatga gtacaagctg
     1321 gcgcaagccc ttcatgagat gagagagcaa catgatgccc aagtgaggct gtataaggag
     1381 gagctggagc agacttacca tgccaaactt gagaatgcca gactgtcatc agagatgaat
     1441 acttctactg tcaacagtgc cagggaagaa ctgatggaaa gccgcatgag aattgagagc
     1501 ctttcatccc agctttctaa tctacagaaa gagtctagag catgtttgga aaggattcaa
     1561 gaattagagg acttgcttgc taaagaaaaa gacaactctc gtcgcatgct gacagacaaa
     1621 gagagagaga tggcggaaat aagggatcaa atgcagcaac agctgaatga ctatgaacag
     1681 cttcttgatg taaagttagc cctggacatg gaaatcagtg cttacaggaa actcttagaa
     1741 ggcgaagaag agaggttgaa gctgtctcca agcccttctt cccgtgtgac agtatcccga
     1801 gcatcctcaa gtcgtagtgt acgtacaact agaggaaagc ggaagagggt tgatgtggaa
     1861 gaatcagagg cgagtagtag tgttagcatc tctcattccg cctcagccac tggaaatgtt
     1921 tgcatcgaag aaattgatgt tgatgggaaa tttatccgct tgaagaacac ttctgaacag
     1981 gatcaaccaa tgggaggctg ggagatgatc agaaaaattg gagacacatc agtcagttat
     2041 aaatatacct caagatatgt gctgaaggca ggccagactg ttacaatttg ggctgcaaac
     2101 gctggtgtca cagccagccc cccaactgac ctcatctgga agaaccagaa ctcgtggggc
     2161 actggcgaag atgtgaaggt tatattgaaa aattctcagg gagaggaggt tgctcaaaga
     2221 agtacagtct ttaaaacaac catacctgaa gaagaggagg aggaggaaga agcagctgga
     2281 gtggttgttg aggaagaact tttccaccag cagggaaccc caagagcatc caatagaagc
     2341 tgtgcaatta tgtaatgcta gatcataatc agccatacca catttgtaga ggttttactt
     2401 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2461 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2521 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2581 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2641 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2701 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2761 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2821 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2881 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2941 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3001 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3061 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3181 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3361 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3421 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3541 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3601 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3661 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3721 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3781 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3841 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3901 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3961 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4021 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4081 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4141 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4201 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4261 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4321 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4381 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4441 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4501 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4561 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4621 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4681 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4741 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4801 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4861 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4921 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4981 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5041 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5101 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5161 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5221 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5281 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5341 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5401 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5461 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5521 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5581 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5641 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     5701 tacggggtca ttagttcata gcccatatat ggagttccg
//
