LOCUS       sequence-220569-        5360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of vimentin with C-terminal Spot-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5360)
  TITLE     ChromoTek Spot-tag plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5360)
  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            "Unpublished"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: pVimentin-Spot-Tag
FEATURES             Location/Qualifiers
     source          1..5360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1989..2024
                     /codon_start=1
                     /product="engineered Spot-Tag(R) peptide that binds with 
                     high affinity to a nanobody"
                     /label=Spot-Tag
                     /translation="PDRVRAVSHWSS"
     polyA_signal    2136..2257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2173..2192)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2227..2246
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2264..2719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2613..2632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2746..2850
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2852..3209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2852..3043
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2875..2895)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3060..3195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3244..4038
                     /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             3244..4038
                     /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(3298..3317)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3908..3927
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4226..4245)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4270..4317
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4646..5234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5135..5154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgccacc atgtccacca
      601 ggtccgtgtc ctcgtcctcc taccgcagga tgttcggcgg cccgggcacc gcgagccggc
      661 cgagctccag ccggagctac gtgactacgt ccacccgcac ctacagcctg ggcagcgcgc
      721 tgcgccccag caccagccgc agcctctacg cctcgtcccc gggcggcgtg tatgccacgc
      781 gctcctctgc cgtgcgcctg cggagcagcg tgcccggggt gcggctcctg caggactcgg
      841 tggacttctc gctggccgac gccatcaaca ccgagttcaa gaacacccgc accaacgaga
      901 aggtggagct gcaggagctg aatgaccgct tcgccaacta catcgacaag gtgcgcttcc
      961 tggagcagca gaataagatc ctgctggccg agctcgagca gctcaagggc caaggcaagt
     1021 cgcgcctggg ggacctctac gaggaggaga tgcgggagct gcgccggcag gtggaccagc
     1081 taaccaacga caaagcccgc gtcgaggtgg agcgcgacaa cctggccgag gacatcatgc
     1141 gcctccggga gaaattgcag gaggagatgc ttcagagaga ggaagccgaa aacaccctgc
     1201 aatctttcag acaggatgtt gacaatgcgt ctctggcacg tcttgacctt gaacgcaaag
     1261 tggaatcttt gcaagaagag attgcctttt tgaagaaact ccacgaagag gaaatccagg
     1321 agctgcaggc tcagattcag gaacagcatg tccaaatcga tgtggatgtt tccaagcctg
     1381 acctcacggc tgccctgcgt gacgtacgtc agcaatatga aagtgtggct gccaagaacc
     1441 tgcaggaggc agaagaatgg tacaaatcca agtttgctga cctctctgag gctgccaacc
     1501 ggaacaatga cgccctgcgc caggcaaagc aggagtccac tgagtaccgg agacaggtgc
     1561 agtccctcac ctgtgaagtg gatgccctta aaggaaccaa tgagtccctg gaacgccaga
     1621 tgcgtgaaat ggaagagaac tttgccgttg aagctgctaa ctaccaagac actattggcc
     1681 gcctgcagga tgagattcag aatatgaagg aggaaatggc tcgtcacctt cgtgaatacc
     1741 aagacctgct caatgttaag atggcccttg acattgagat tgccacctac aggaagctgc
     1801 tggaaggcga ggagagcagg atttctctgc ctcttccaaa cttttcctcc ctgaacctga
     1861 gggaaactaa tctggattca ctccctctgg ttgataccca ctcaaaaagg acacttctga
     1921 ttaagacggt tgaaactaga gatggacagg ttatcaacga aacttctcag catcacgatg
     1981 accttgaacc agaccgcgtg cgcgccgtga gccattggag cagctgactg atcataatca
     2041 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2101 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2161 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2221 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2281 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2341 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2401 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2461 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2521 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2581 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2641 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2701 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2761 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2821 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2881 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2941 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3001 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3061 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3121 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3181 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3241 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3301 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3361 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3421 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3481 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3541 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3601 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3661 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3721 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3781 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3841 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3901 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3961 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4021 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4081 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4141 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4201 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4261 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4321 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4381 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4441 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5341 ccatgcatta gttattaata
//
