LOCUS       Exported                6510 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  APEX2-Lamin B1 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP - FLAG-APEX2- hs Lamin B1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  TITLE     APEX2-hsLamin B1, bioRxiv - The versatility of Ascorbate Peroxidase 
            in mapping nuclear lamina interactions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6510
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     144..164
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      232..241
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             241..264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2995..3116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3032..3051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3086..3105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3123..3578)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3260..3281)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3472..3491
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3605..3709
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3734..3754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3778..3988
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3800..3996
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3847..3982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3909..3928
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4031..4825
                     /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(4085..4104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4695..4714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5013..5032)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5057..5104
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5433..6021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5922..5941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6196..6499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6500..193
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
       61 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      121 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      181 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
      241 gactacaagg atgacgacga taaggggaaa tcatacccaa cagtgtccgc agactaccag
      301 gatgccgtgg agaaagccaa gaagaagctg agagggttta tcgcagaaaa gaggtgcgca
      361 cctctgatgc tgagactggc tttccacagc gcaggcacct ttgacaaggg aaccaaaaca
      421 ggcggaccct ttggaacaat caagcaccct gctgaactgg cacatagtgc caacaatggg
      481 ctggacatcg ccgtgcggct gctggaacct ctgaaagcag agttcccaat tctgtcctac
      541 gccgattttt atcagctggc aggagtggtc gctgtggagg tcactggggg ccccaaggtg
      601 cctttccacc caggacggga ggacaaacca gaaccacctc cagaggggcg cctgccagat
      661 ccgacaaagg gctccgacca tctgcgagat gtgtttggga aagctatggg cctgactgac
      721 caggatatcg tcgcactgtc tggagggcac accattggcg ccgctcataa ggaaaggtca
      781 ggcttcgagg gaccctggac aagcaacccc ctgattttcg acaattctta ctttactgaa
      841 ctgctgagtg gagagaagga agggctgctg cagctgccca gcgacaaagc cctgctgtcc
      901 gatcccgtgt tcagacctct ggtcgataag tatgcagccg acgaggatgc ttttttcgca
      961 gattacgcag aagcacatca gaagctgtca gaactgggat ttgccgacgc caagggctcg
     1021 ggctcgacct cgggctcggg ctccggactc agatctcgag gaggtggggg atctggtggc
     1081 ggaggttcag gaggtggagg atcgatggcg actgcgaccc ccgtgccgcc gcggatgggc
     1141 agccgcgctg gcggccccac cacgccgctg agccccacgc gcctgtcgcg gctccaggag
     1201 aaggaggagc tgcgcgagct caatgaccgg ctggcggtgt acatcgacaa ggtgcgcagc
     1261 ctggagacgg agaacagcgc gctgcagctg caggtgacgg agcgcgagga ggtgcgcggc
     1321 cgtgagctca ccggcctcaa ggcgctctac gagaccgagc tggccgacgc gcgacgcgcg
     1381 ctcgacgaca cggcccgcga gcgcgccaag ctgcagatcg agctgggcaa gtgcaaggcg
     1441 gaacacgacc agctgctcct caactatgct aagaaggaat ctgatcttaa tggcgcccag
     1501 atcaagcttc gagaatatga agcagcactg aattcgaaag atgcagctct tgctactgca
     1561 cttggtgaca aaaaaagttt agagggagat ttggaggatc tgaaggatca gattgcccag
     1621 ttggaagcct ccttagctgc agccaaaaaa cagttagcag atgaaacttt acttaaagta
     1681 gatttggaga atcgttgtca gagccttact gaggacttgg agtttcgcaa aagcatgtat
     1741 gaagaggaga ttaacgagac cagaaggaag catgaaacgc gcttggtaga ggtggattct
     1801 gggcgtcaaa ttgagtatga gtacaagctg gcgcaagccc ttcatgagat gagagagcaa
     1861 catgatgccc aagtgaggct gtataaggag gagctggagc agacttacca tgccaaactt
     1921 gagaatgcca gactgtcatc agagatgaat acttctactg tcaacagtgc cagggaagaa
     1981 ctgatggaaa gccgcatgag aattgagagc ctttcatccc agctttctaa tctacagaaa
     2041 gagtctagag catgtttgga aaggattcaa gaattagagg acttgcttgc taaagaaaaa
     2101 gacaactctc gtcgcatgct gacagacaaa gagagagaga tggcggaaat aagggatcaa
     2161 atgcagcaac agctgaatga ctatgaacag cttcttgatg taaagttagc cctggacatg
     2221 gaaatcagtg cttacaggaa actcttagaa ggcgaagaag agaggttgaa gctgtctcca
     2281 agcccttctt cccgtgtgac agtatcccga gcatcctcaa gtcgtagtgt acgtacaact
     2341 agaggaaagc ggaagagggt tgatgtggaa gaatcagagg cgagtagtag tgttagcatc
     2401 tctcattccg cctcagccac tggaaatgtt tgcatcgaag aaattgatgt tgatgggaaa
     2461 tttatccgct tgaagaacac ttctgaacag gatcaaccaa tgggaggctg ggagatgatc
     2521 agaaaaattg gagacacatc agtcagttat aaatatacct caagatatgt gctgaaggca
     2581 ggccagactg ttacaatttg ggctgcaaac gctggtgtca cagccagccc cccaactgac
     2641 ctcatctgga agaaccagaa ctcgtggggc actggcgaag atgtgaaggt tatattgaaa
     2701 aattctcagg gagaggaggt tgctcaaaga agtacagtct ttaaaacaac catacctgaa
     2761 gaagaggagg aggaggaaga agcagctgga gtggttgttg aggaagaact tttccaccag
     2821 cagggaaccc caagagcatc caatagaagc tgtgcaatta tgtaaggatc caccggatct
     2881 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2941 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3121 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3181 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3241 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3301 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3361 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3421 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3481 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3541 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3661 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3721 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3901 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3961 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4021 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4081 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4141 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4201 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4261 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4321 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4381 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4441 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4501 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4561 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4621 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4681 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4741 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4801 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4861 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4921 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4981 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5041 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5101 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5161 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5221 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5281 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5341 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5401 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5461 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5521 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5581 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5641 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5701 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5761 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5821 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5881 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5941 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6001 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6061 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6121 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     6181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6481 agtcatcgct attaccatgg tgatgcggtt
//
