LOCUS       sequence-134190-        7273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone for human Fibrillarin with C-terminal 3xFLAG-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7273)
  AUTHORS   Stamm
  TITLE     p3xFLAG_Fibrillarin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7273)
  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            "Unpublished"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: pCMV14_Fibrillarin
FEATURES             Location/Qualifiers
     source          1..7273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1749..1760)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             1956..2021
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2032..2654
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2231..2250)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2678..2698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2696..2993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3047..3841
                     /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(3101..3120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3711..3730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4493..4627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4530..4549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4584..4603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4632..4651)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4633..4651)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4665..4681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4665..4681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4678..4700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4689..4705
                     /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(4713..4719,4720..4737,4738..4743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4758..4779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4896..4913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5067..5655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5147..5166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5826..6617,6618..6686))
                     /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     6449..6468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6687..6791)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6818..7246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6905..6924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7115..7136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
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 attaagcttg cggccgcgaa ttcatcgata gatctgatat
      961 cggtaccagt cgacatgaag ccaggattca gtccccgtgg gggtggcttt ggcggccgag
     1021 ggggctttgg tgaccgtggt ggtcgtggag gccgaggggg ctttggcggg ggccgaggtc
     1081 gaggcggagg ctttagaggt cgtggacgag gaggaggtgg aggcggcggc ggcggtggag
     1141 gaggaggaag aggtggtgga ggcttccatt ctggtggcaa ccggggtcgt ggtcggggag
     1201 gaaaaagagg aaaccagtcg gggaagaatg tgatggtgga gccgcatcgg catgagggtg
     1261 tcttcatttg tcgaggaaag gaagatgcac tggtcaccaa gaacctggtc cctggggaat
     1321 cagtttatgg agagaagaga gtctcgattt cggaaggaga tgacaaaatt gagtaccgag
     1381 cctggaaccc cttccgctcc aagctagcag cagcaatcct gggtggtgtg gaccagatcc
     1441 acatcaaacc gggggctaag gttctctacc tcggggctgc ctcgggcacc acggtctccc
     1501 atgtctctga catcgttggt ccggatggtc tagtctatgc agtcgagttc tcccaccgct
     1561 ctggccgtga cctcattaac ttggccaaga agaggaccaa catcattcct gtgatcgagg
     1621 atgctcgaca cccacacaaa taccgcatgc tcatcgcaat ggtggatgtg atctttgctg
     1681 atgtggccca gccagaccag acccggattg tggccctgaa tgcccacacc ttcctgcgta
     1741 atggaggaca ctttgtgatt tccattaagg ccaactgcat tgactccaca gcctcagccg
     1801 aggccgtgtt tgcctccgaa gtgaaaaaga tgcaacagga gaacatgaag ccgcaggagc
     1861 agttgaccct tgagccatat gaaagagacc atgccgtggt cgtgggagtg tacaggccac
     1921 cccccaaggt gaagaactct agaggatccc gggctgacta caaagaccat gacggtgatt
     1981 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgatcc cgggtggcat
     2041 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2101 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2161 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2221 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2281 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2341 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     2401 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     2461 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     2521 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     2581 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     2641 atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt
     2701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2761 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2821 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2881 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2941 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggaatt
     3001 gatcagcttg ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3061 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3121 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3181 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3241 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3301 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3361 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3421 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3481 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3541 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3601 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3661 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3721 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3781 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3841 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3901 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3961 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc
     4021 gatcccctcg cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac
     4081 cggcagtgca aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc
     4141 cccgaactgc aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc
     4201 ctgcgcctga tacagaacga attgcttgca ggcatctcat gagtgtgtct tcccgttttc
     4261 cgcctgaggt cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc gagctgctca
     4321 ccacccactc gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4381 tccgccgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     4441 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4501 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4561 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4621 ctggatcaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct
     4681 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4741 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4861 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5041 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5101 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5161 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5221 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5281 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5341 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5401 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5461 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5521 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5581 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5641 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5701 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5761 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5821 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5881 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5941 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6001 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6061 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6121 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6181 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6241 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6301 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6361 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6421 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6481 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6541 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6601 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6661 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6721 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6781 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     6841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6961 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7021 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7141 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7261 acgcttacaa ttt
//
