LOCUS       Exported                6299 bp DNA     circular SYN 10-SEP-2025
DEFINITION  Vector with a neomycin resistance marker for expression of 
            N-terminally 3xFLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-CMV-10
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6299
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             928..930
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /translation="M"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG epitope tags, followed by an 
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    994..1060
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1058..1680
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1722..2019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2073..2867
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3519..3653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3691..3707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4093..4681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4852..5712)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5713..5817)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5844..6299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata
     1021 gatctgatat cggtaccagt cgactctaga ggatcccggg tggcatccct gtgacccctc
     1081 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     1141 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     1201 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     1261 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     1321 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     1381 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     1441 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     1501 acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca
     1561 ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag
     1621 ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt
     1681 gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1861 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1921 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     1981 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggaattgatc agcttgggat
     2041 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     2101 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2161 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2221 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2281 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2341 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2401 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2461 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2521 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2581 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2641 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2701 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2761 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2821 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     2881 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     2941 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3001 ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggctcgatc ccctcgcgag
     3061 ttggttcagc tgctgcctga ggctggacga cctcgcggag ttctaccggc agtgcaaatc
     3121 cgtcggcatc caggaaacca gcagcggcta tccgcgcatc catgcccccg aactgcagga
     3181 gtggggaggc acgatggccg ctttggtcga cccggacggg acgctcctgc gcctgataca
     3241 gaacgaattg cttgcaggca tctcatgagt gtgtcttccc gttttccgcc tgaggtcact
     3301 gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc tgctcaccac ccactcgcca
     3361 agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg ccgatcataa
     3421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaattccc
     3661 tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     3721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5821 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//
