LOCUS       sequence-356071-        7471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV overexpression of SNAP-tag fast fusion to Lamin B1 for nuclear 
            envelope localization in mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Frei MS, Koch B, Hiblot J, Johnsson K
  TITLE     Live-Cell Fluorescence Lifetime Multiplexing Using Synthetic 
            Fluorescent Probes.
  JOURNAL   ACS Chem Biol. 2022 Jun 17;17(6):1321-1327. doi: 
            10.1021/acschembio.2c00041. Epub 2022 May 18.
  PUBMED    35584304
REFERENCE   2  (bases 1 to 7471)
  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"; doi: 
            "10.1021/acschembio.2c00041"; journalName: "ACS Chem Biol"; date: 
            "2022-06-17- 17"; volume: "17"; issue: "6"; pages: "1321-1327"
COMMENT     Sequence Label: pcDNA5/FRT/TO_STf_LamB1
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3423..3440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3429..3653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3786..3805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3937..3984
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3992..5014
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3992..5014
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5144..5265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5181..5200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5235..5254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5314..5330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5314..5330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5327..5349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5338..5354
                     /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(5362..5368,5369..5386,5387..5392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5407..5428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5545..5562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5716..6304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5796..5815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6475..7266,7267..7335))
                     /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     7098..7117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7336..7440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcaa ccatggataa agactgcgaa atgaaacgca ctaccctgga
     1081 cagcccgctg ggtaaactgg aactgagcgg ttgcgagcag ggcctgcacc gcatcatttt
     1141 cctgggcaaa ggtacgagcg ctgcggatgc ggtcgaagta ccggctccgg ctgcggttct
     1201 gggtggcccg gagccgctga tgcaggctac cgcatggctg aacgcatatt ttcaccagcc
     1261 agaggcgatt gaagagttcc ctgtaccggc cctgcatcac ccggtatttc agcaggaatc
     1321 ctttacccgt caggtactgt ggaaactgct gaaagtagtg aaattcggcg aggtgatctc
     1381 ctattcccat ctggcggcgc tggctggcaa cccagcggca actgcggcgg tgaaaaccgc
     1441 tctgtctggt aatccggtgc caattctgat cccgtgccac cgtgtggttc agggcgacct
     1501 ggacgtaggt ggctacgaag gcggcctggc tgtgaaagaa tggctgctgg cgcacgaagg
     1561 tcaccgtctg ggtaaaccag gcctgggctc cggactcaga tctcgagctc aagcttcgat
     1621 ggcgactgcg acccccgtgc cgccgcggat gggcagccgc gctggcggcc ccaccacgcc
     1681 gctgagcccc acgcgcctgt cgcggctcca ggagaaggag gagctgcgcg agctcaatga
     1741 ccggctggcg gtgtacatcg acaaggtgcg cagcctggag acggagaaca gcgcgctgca
     1801 gctgcaggtg acggagcgcg aggaggtgcg cggccgtgag ctcaccggcc tcaaggcgct
     1861 ctacgagacc gagctggccg acgcgcgacg cgcgctcgac gacacggccc gcgagcgcgc
     1921 caagctgcag atcgagctgg gcaagtgcaa ggcggaacac gaccagctgc tcctcaacta
     1981 tgctaagaag gaatctgatc ttaatggcgc ccagatcaag cttcgagaat atgaagcagc
     2041 actcaattcg aaagatgcag ctcttgctac tgcacttggt gacaaaaaaa gtttagaggg
     2101 agatttggag gatctgaagg atcagattgc ccagttggaa gcctccttag ctgcagccaa
     2161 aaaacagtta gcagatgaaa ctttacttaa agtagatttg gagaatcgtt gtcagagcct
     2221 tactgaggac ttggagtttc gcaaaagcat gtatgaagag gagattaacg agaccagaag
     2281 gaagcatgaa acgcgcttgg tagaggtgga ttctgggcgt caaattgagt atgagtacaa
     2341 gctggcgcaa gcccttcatg agatgagaga gcaacatgat gcccaagtga ggctgtataa
     2401 ggaggagctg gagcagactt accatgccaa acttgagaat gccagactgt catcagagat
     2461 gaatacttct actgtcaaca gtgccaggga agaactgatg gaaagccgca tgagaattga
     2521 gagcctttca tcccagcttt ctaatctaca gaaagagtct agagcatgtt tggaaaggat
     2581 tcaagaatta gaggacttgc ttgctaaaga aaaagacaac tctcgtcgca tgctgacaga
     2641 caaagagaga gagatggcgg aaataaggga tcaaatgcag caacagctga atgactatga
     2701 acagcttctt gatgtaaagt tagccctgga catggaaatc agtgcttaca ggaaactctt
     2761 agaaggcgaa gaagagaggt tgaagctgtc tccaagccct tcttcccgtg tgacagtatc
     2821 ccgagcatcc tcaagtcgta gtgtacgtac aactagagga aagcggaaga gggttgatgt
     2881 ggaagaatca gaggcgagta gtagtgttag catctctcat tccgcctcag ccactggaaa
     2941 tgtttgcatc gaagaaattg atgttgatgg gaaatttatc cgcttgaaga acacttctga
     3001 acaggatcaa ccaatgggag gctgggagat gatcagaaaa attggagaca catcagtcag
     3061 ttataaatat acctcaagat atgtgctgaa ggcaggccag actgttacaa tttgggctgc
     3121 aaacgctggt gtcacagcca gccccccaac tgacctcatc tggaagaacc agaactcgtg
     3181 gggcactggc gaagatgtga aggttatatt gaaaaattct cagggagagg aggttgctca
     3241 aagaagtaca gtctttaaaa caaccatacc tgaagaagag gaggaggagg aagaagcagc
     3301 tggagtggtt gttgaggaag aacttttcca ccagcaggga accccaagag catccaatag
     3361 aagctgtgca attatgtagt aaccgctcga gtctagaggg cccgtttaaa cccgctgatc
     3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3661 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3841 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3901 caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta
     3961 ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt
     4021 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4081 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4141 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     4201 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     4261 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4321 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     4381 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4441 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4501 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4561 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4621 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4681 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4741 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     4801 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4861 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4921 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4981 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
     5041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5281 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7441 cacatttccc cgaaaagtgc cacctgacgt c
//
