LOCUS       sequence-194147-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SNAP-Tag and CaaX box (Farnesylation Signal) in 
            Mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5418)
  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            "Unpublished"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: pSNAP-CaaX Control Plasmid
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..286
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     243..263
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(449..1240,1241..1309))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1072..1091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1310..1401)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1420..1447)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     polyA_signal    1475..1596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1486..1505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1540..1559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1870..1889
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1870..1888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1979..2524)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     promoter        complement(2576..2594)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2640..2843)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2669..2689)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2844..3147
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3322..3910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3402..3421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4239..4286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4311..4330
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4518..5312)
                     /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             complement(4518..5312)
                     /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(4629..4648)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5239..5258
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5347..5418,1..286))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5361..5418,1..78)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
       61 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
      121 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acaccctaac tgacacacat tccacagctg gttctttccg cctcaggatt cttccttttt
      301 caatgtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      361 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      421 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      481 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      541 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      601 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      661 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      721 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      781 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      841 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
      901 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      961 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1021 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     1081 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1141 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1201 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1261 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1321 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1381 gtatttagaa aaataaacaa aagagtttgt agaaacgcaa aaagaccatc cgtcaggatg
     1441 gccttctgct tattaacgct tacaatttac gcgttaagat acattgatga gtttggacaa
     1501 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1561 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1621 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1681 tgtggtatgg ctgattatga tcatgaacag actgtgagga ctgaggggcc tgaaatgagc
     1741 cttgggactg tgaatctaaa atacacaaac aattagaatc agtagtttaa cacattatac
     1801 acttaaaaat tggatctcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1861 agtgaattgt aatacgactc actatagggc gaattgggta cacttacctg gtaccgggcc
     1921 ccccctcgag tcaggatcct caggagagca cacacttgca gctcatgcag cctgcaggac
     1981 ccagcccagg cttgcccagt ctgtggccct cgtgggccag cagccactct ttcacggcga
     2041 gcccgccctc gtagcccccc acggcgccag agctagacac cacccggtgg caggggatca
     2101 gaatgggcac gggatttccg ctcagggcgg ttttcacggc ggcggtggcg gcgggattgc
     2161 cggccagggc ggccagctgc tggtagctga tgacctctcc gaacttcacc actttcagca
     2221 gtttccacag cacctggcgg gtaaagctct cctgctggaa cactgggtgg tgcagggctg
     2281 gcacagggaa ctcctcgatg gcctcaggct ggtgaaagta ggcgttgagc caggcggtgg
     2341 cctgcatcag tggctctggt ccgcccagca cggcggctgg ggcaggcact tccacggcgt
     2401 cggcggcaga tgttcctttg cccagcagct tgatctcgtg caggccctgt tcgcacccag
     2461 acagttccag cttgcccaga gggctatcca gggtggtgcg cttcatttcg cagtctttgt
     2521 ccatggtaag cttgatatcg cggccgccac cgcggtggag ctccagcttt tgttcccttt
     2581 agtgagggtt aatttcgagc ttggcgtaat cgctagcgga tctgacggtt cactaaacca
     2641 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     2701 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     2761 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     2821 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     2881 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     2941 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     3001 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     3061 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     3121 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     3181 cccgtaattg attactatta ataactaatg catggcggta atacggttat ccacagaatc
     3241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     4081 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     4141 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     4201 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     4261 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     4321 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     4381 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     4441 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     4501 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4561 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagt
     4621 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4681 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     4741 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     4801 aacagttcgg ctggcgcgag cccctgatgt tcttcgtcca gatcatcctg atcgacaaga
     4861 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     4921 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     4981 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5041 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5101 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     5161 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5221 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5281 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5341 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5401 agaggccgag gcggcctc
//
