LOCUS       sequence-355855-        5515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty plasmid containing Flag tag for C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5515)
  AUTHORS   Cerqua M, Botti O, Arigoni M, Gioelli N, Serini G, Calogero R, 
            Boccaccio C, Comoglio PM, Altintas DM
  TITLE     MET14 promotes a ligand-dependent, AKT-driven invasive growth.
  JOURNAL   Life Sci Alliance. 2022 May 30;5(10). pii: 5/10/e202201409. doi: 
            10.26508/lsa.202201409. Print 2022 Oct.
  PUBMED    35636967
REFERENCE   2  (bases 1 to 5515)
  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: "Life Sci 
            Alliance."; date: "2022-05-30"; volume: "5(10). pii: 
            5/10/e202201409. doi"; pages: " 10.26508/lsa.202201409. Print 2022 
            Oct"
COMMENT     Sequence Label: pCDNA3.1_3XFlag
FEATURES             Location/Qualifiers
     source          1..5515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(90..109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        281..660
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        661..864
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     815..835
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     909..928
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        909..927
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             978..1055
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKGDYKDDDDKIDYKDDDDK"
     primer_bind     complement(1152..1169)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1158..1382
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1428..1856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1515..1534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1725..1746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1865..1885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1870..2199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2050..2185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2112..2131
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2266..3060
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2320..2339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2930..2949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3234..3355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3325..3344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3404..3420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3404..3420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3417..3439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3428..3444
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3452..3458,3459..3476,3477..3482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3497..3518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3635..3652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3806..4394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3886..3905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4565..5356,5357..5425))
                     /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"
     primer_bind     5188..5207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5426..5515,1..15))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
       61 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      121 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      181 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      241 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      301 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      361 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      421 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      481 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      541 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      601 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      661 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      721 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      781 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      841 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      901 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      961 ttggcggtgg gggtggcgat tacaaggatg acgacgataa gggagattac aaggatgacg
     1021 acgataagat cgattacaag gatgacgacg ataagtaagg atccactagt ccagtgtggt
     1081 ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     1141 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1201 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1261 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1321 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1381 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1441 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1501 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1561 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1621 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1681 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1741 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1801 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1861 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1921 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1981 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2041 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2101 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2161 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2221 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2281 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2341 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2401 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2461 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2521 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2581 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2641 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2701 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2761 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     2821 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2881 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2941 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3001 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3061 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3121 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3181 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3361 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3421 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3481 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaaca
//
