LOCUS       sequence-212885-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresssion of Flag-tagged NF-kB p49.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  TITLE     Georg Weber Lab Plasmids
REFERENCE   2  (bases 1 to 6538)
  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"
COMMENT     Sequence Label: pFLAG-p49
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             738..761
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2276..2293)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2282..2506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2638..3226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2718..2737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3555..3602
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3627..3646
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3834..4628)
                     /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(3834..4628)
                     /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(3945..3964)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4555..4574
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4663..5020)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4677..4812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4829..5020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4977..4997
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5080..5871,5872..5940))
                     /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     5703..5722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5941..6045)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6072..6527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6159..6178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6369..6390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcggctctg cagcatggac tacaaggacg acgatgacaa ggagagttgc tacaacccag
      781 gtctggatgg tattattgaa tatgatgatt tcaaattgaa ctcctccatt gtggaaccca
      841 aggagccagc cccagaaaca gctgatggcc cctacctggt gatcgtggaa cagcctaagc
      901 agagaggctt ccgatttcga tatggctgtg aaggcccctc ccatggagga ctgcccggtg
      961 cctccagtga gaagggccga aagacctatc ccactgtcaa gatctgtaac tacgagggac
     1021 cagccaagat cgaggtggac ctggtaacac acagtgaccc acctcgtgct catgcccaca
     1081 gtctggtggg caagcaatgc tcggagctgg ggatctgcgc cgtttctgtg gggcccaagg
     1141 acatgactgc ccaatttaac aacctgggtg tcctgcatgt gactaagaag aacatgatgg
     1201 ggactatgat acaaaaactt cagaggcagc ggctccgctc taggccccag ggccttacgg
     1261 aggccgagca gcgggagctg gagcaagagg ccaaagaact gaagaaggtg atggatctga
     1321 gtatagtgcg gctgcgcttc tctgccttcc ttagagccag tgatggctcc ttctccctgc
     1381 ccctgaagcc agtcatctcc cagcccatcc atgacagcaa atctccgggg gcatcaaacc
     1441 tgaagatttc tcgaatggac aagacagcag gctctgtgcg gggtggagat gaagtttatc
     1501 tgctttgtga caaggtgcag aaagatgaca ttgaggttcg gttctatgag gatgatgaga
     1561 atggatggca ggcctttggg gacttctctc ccacagatgt gcataaacag tatgccattg
     1621 tgttccggac acccccctat cacaagatga agattgagcg gcctgtaaca gtgtttctgc
     1681 aactgaaacg caagcgagga ggggacgtgt ctgattccaa acagttcacc tattaccctc
     1741 tggtggaaga caaggaagag gtgcagcgga agcggaggaa ggccttgccc accttctccc
     1801 agcccttcgg gggtggctcc cacatgggtg gaggctctgg gggtgcagcc gggggctacg
     1861 gaggagctgg aggaggtgag ggggtactga tggagggagg ggtaaaggta agagaagctg
     1921 tggaggaaaa aaatctgggg gaggccgggc gtggtggctc acgcctgtaa tccagccctt
     1981 tgggaggcca aggcaggcag attacctgag atcaggagtt caagaccagc ttggccaaca
     2041 gcgtgaaacc tcgtctctac taaaaataca aacattagct gggcatggtg gcaggcgcct
     2101 gtaatcccag ctactcggga ggctgaggca ggagaatcgc ttgaaccctg ggagacaaga
     2161 ggttgcagta agctgagatc acaccactgc actccaggct gagccgaatt ctgcagatat
     2221 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2281 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2341 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2401 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2461 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2521 agaaccagtg gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3361 aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc
     3421 tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg
     3481 ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     3541 aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc
     3601 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct
     3661 agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     3721 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     3781 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
     3841 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     3901 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     3961 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     4021 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     4081 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     4141 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     4201 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     4261 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     4321 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     4381 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     4441 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     4501 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     4561 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     4621 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc
     4681 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     4741 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4801 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     4861 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     4921 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     4981 taactgacac acattccaca gctggttctt tccgcctcag gactcttcct ttttcaataa
     5041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6061 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6121 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6181 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6241 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6301 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6361 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6421 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6481 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgcgcgtt
//
