LOCUS       sequence-439384-        6442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Tagln2-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6442)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 6442)
  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: Tagln2-GFP
FEATURES             Location/Qualifiers
     source          1..6442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(134..181)
                     /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)."
     primer_bind     313..332
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    465..689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     678..695
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(739..1452,1453..1455,1456..1458))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     1133..1152
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1392..1413
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1471..1491)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2196..2220)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2230..2248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2251..2269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(2271..2474)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2300..2320)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2475..2854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3026..3045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3120..3224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3225..3293,3294..4085)
                     /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     complement(3443..3462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4256..4844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4745..4764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4998..5015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5132..5153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5168..5173,5174..5191,5192..5198)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5206..5222
                     /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)."
     primer_bind     5211..5233
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5230..5246
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5230..5246
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5295..5416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5306..5325)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5360..5379
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5546..6442,1..126))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(join(5546..6442,1..126))
                     /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
                     "
ORIGIN
        1 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
       61 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
      121 ctttttggcc aaggaagttc ctatactttc tagagaatag gaacttcgga ataggaactt
      181 ctaggtacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
      241 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
      301 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
      361 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg
      421 gggatacccc ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc
      481 atccccagca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
      541 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
      601 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
      661 aacagatggc tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc
      721 tctagactcg agtcactatt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
      781 ggtcacgaac tccttaagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
      841 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
      901 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
      961 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     1021 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1081 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1141 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1201 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     1261 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     1321 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1381 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1441 ctcgcccttg ctcaccatgg tggtaccttc accttgaaaa tataaatttt cgcggccgcg
     1501 gaggatctgg cgtggcatcc cgtagccagt catgcctgcc tgagacgccc cgcggttggt
     1561 gcccatctgt aacccgatca cgttcttgcc ctcttgcagc tggttatccg agaagttccg
     1621 aggattctcc ttggatttct tagggaacca gttgggatcc ccagagaaga gcccatcatc
     1681 tcgggctact gccagcccac ccagattcat cagcgtccgc tgcacacagg ccatgttctt
     1741 tccttcccag aggtccacag tttggaagat gtcagtggtg ttaatgccat agcgctcagc
     1801 tgcttgcagg aactgagaga tctgctccat ctgcttgaag gccatggtgg aggcctggat
     1861 cttctttact ggggcctgcc cctcggggta ctgtgcatta atgagctcac atagcaccgt
     1921 gccatccttg agccagttct ggaagttctc gcgtccaggc tggggccggc ccacatcctt
     1981 tcggcactgg gtggtgatcc actggatcag gatctgctcc agatctgcat catattgttt
     2041 ctcaatcttc tgctgcacct cccggctcag gccatatgca ggtcccctgt tggccatgca
     2101 ggcctaagct taagtttaaa cgctagagtc cggaggctgg atcggtcccg gtgtcttcta
     2161 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     2221 cgtcgacgat ctctatcact gatagggaga tctctatcac tgatagggag agctctgctt
     2281 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     2341 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     2401 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     2461 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     2521 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     2581 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     2641 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     2701 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     2761 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     2821 gattactatt aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg
     2881 cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct
     2941 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     3001 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3061 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     3121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3181 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3241 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3301 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3361 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3421 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3481 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3541 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3601 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3661 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3721 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3781 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3841 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3901 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3961 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4021 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4081 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4141 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4201 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4261 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4321 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4381 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4441 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4501 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4561 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4621 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4681 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4741 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4801 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4861 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4921 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4981 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     5041 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5101 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     5161 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5221 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     5281 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     5341 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5401 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     5461 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     5521 tggaatcgaa atctcgtagt acgtgctatt cctttgccct cggacgagtg ctggggcgtc
     5581 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
     5641 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
     5701 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
     5761 gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc
     5821 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
     5881 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
     5941 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
     6001 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
     6061 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
     6121 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
     6181 tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa
     6241 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
     6301 tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg
     6361 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
     6421 ttacccgcag gacatatcca cg
//
