LOCUS       sequence-220574-        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  JOURNAL   Unpublished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            "Unpublished"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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             622..657
                     /codon_start=1
                     /product="engineered Spot-Tag(R) peptide that binds with 
                     high affinity to a nanobody"
                     /label=Spot-Tag
                     /translation="PDRVRAVSHWSS"
     polyA_signal    2527..2648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2564..2583)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2618..2637
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2655..3110)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2792..2813)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3004..3023
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3137..3241
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3243..3600
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3243..3434
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3266..3286)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3451..3586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3635..4429
                     /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             3635..4429
                     /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(3689..3708)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4299..4318
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4617..4636)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4661..4708
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5037..5625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5526..5545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat gcctgatcgc gtgcgcgccg tgagccattg gagcagcgcg
      661 actgcgaccc ccgtgccgcc gcggatgggc agccgcgctg gcggccccac cacgccgctg
      721 agccccacgc gcctgtcgcg gctccaggag aaggaggagc tgcgcgagct caatgaccgg
      781 ctggcggtgt acatcgacaa ggtgcgcagc ctggagacgg agaacagcgc gctgcagctg
      841 caggtgacgg agcgcgagga ggtgcgcggc cgtgagctca ccggcctcaa ggcgctctac
      901 gagaccgagc tggccgacgc gcgacgcgcg ctcgacgaca cggcccgcga gcgcgccaag
      961 ctgcagatcg agctgggcaa gtgcaaggcg gaacacgacc agctgctcct caactatgct
     1021 aagaaggaat ctgatcttaa tggcgcccag atcaagcttc gagaatatga agcagcactg
     1081 aattcgaaag atgcagctct tgctactgca cttggtgaca aaaaaagttt agagggagat
     1141 ttggaggatc tgaaggatca gattgcccag ttggaagcct ccttagctgc agccaaaaaa
     1201 cagttagcag atgaaacttt acttaaagta gatttggaga atcgttgtca gagccttact
     1261 gaggacttgg agtttcgcaa aagcatgtat gaagaggaga ttaacgagac cagaaggaag
     1321 catgaaacgc gcttggtaga ggtggattct gggcgtcaaa ttgagtatga gtacaagctg
     1381 gcgcaagccc ttcatgagat gagagagcaa catgatgccc aagtgaggct gtataaggag
     1441 gagctggagc agacttacca tgccaaactt gagaatgcca gactgtcatc agagatgaat
     1501 acttctactg tcaacagtgc cagggaagaa ctgatggaaa gccgcatgag aattgagagc
     1561 ctttcatccc agctttctaa tctacagaaa gagtctagag catgtttgga aaggattcaa
     1621 gaattagagg acttgcttgc taaagaaaaa gacaactctc gtcgcatgct gacagacaaa
     1681 gagagagaga tggcggaaat aagggatcaa atgcagcaac agctgaatga ctatgaacag
     1741 cttcttgatg taaagttagc cctggacatg gaaatcagtg cttacaggaa actcttagaa
     1801 ggcgaagaag agaggttgaa gctgtctcca agcccttctt cccgtgtgac agtatcccga
     1861 gcatcctcaa gtcgtagtgt acgtacaact agaggaaagc ggaagagggt tgatgtggaa
     1921 gaatcagagg cgagtagtag tgttagcatc tctcattccg cctcagccac tggaaatgtt
     1981 tgcatcgaag aaattgatgt tgatgggaaa tttatccgct tgaagaacac ttctgaacag
     2041 gatcaaccaa tgggaggctg ggagatgatc agaaaaattg gagacacatc agtcagttat
     2101 aaatatacct caagatatgt gctgaaggca ggccagactg ttacaatttg ggctgcaaac
     2161 gctggtgtca cagccagccc cccaactgac ctcatctgga agaaccagaa ctcgtggggc
     2221 actggcgaag atgtgaaggt tatattgaaa aattctcagg gagaggaggt tgctcaaaga
     2281 agtacagtct ttaaaacaac catacctgaa gaagaggagg aggaggaaga agcagctgga
     2341 gtggttgttg aggaagaact tttccaccag cagggaaccc caagagcatc caatagaagc
     2401 tgtgcaatta tgtaatgcta gatcataatc agccatacca catttgtaga ggttttactt
     2461 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2521 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2581 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2641 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2701 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2761 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2821 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2881 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2941 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3001 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3061 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3241 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3361 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3421 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3481 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3541 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3601 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3661 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3721 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3781 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3841 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3901 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3961 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4021 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4081 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4141 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4201 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4261 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4321 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4381 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4441 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4501 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4561 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4621 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4681 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4741 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4801 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4861 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4921 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4981 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5041 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5101 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5161 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5221 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5281 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5341 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5401 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5461 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5521 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5581 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5641 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5701 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
