LOCUS       sequence-254949-        4764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses td-iFAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4764)
  AUTHORS   Tebo AG, Pimenta FM, Zhang Y, Gautier A
  TITLE     Improved Chemical-Genetic Fluorescent Markers for Live Cell 
            Microscopy.
  JOURNAL   Biochemistry. 2018 Oct 2;57(39):5648-5653. doi: 
            10.1021/acs.biochem.8b00649. Epub 2018 Sep 17.
  PUBMED    30204425
REFERENCE   2  (bases 1 to 4764)
  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/acs.biochem.8b00649"; journalName: "Biochemistry"; date: 
            "2018-10-2- 2"; volume: "57"; issue: "39"; pages: "5648-5653"
COMMENT     Sequence Label: pAG328 td-iFAST
FEATURES             Location/Qualifiers
     source          1..4764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1387..1416
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1544..1665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1581..1600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1635..1654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1672..2127)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1809..1830)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2021..2040
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2154..2258
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2260..2617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2260..2451
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2283..2303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2468..2603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2652..3446
                     /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             2652..3446
                     /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(2706..2725)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3316..3335
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3634..3653)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3678..3725
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4054..4642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4543..4562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      601 agatctgcca ccatggaaca tgtcgccttc ggcagcgagg atatcgagaa cactctggct
      661 aagatggatg acgggcaact ggacggcttg gcattcgggg ccattcaact ggacggggac
      721 ggtaatattc tccagtacaa tgcagctgaa ggtgacataa ctggacgtga tcctaaacaa
      781 gtgattggca agaatttctt caaagatgtt gctcccggca ccgattcacc tgagttttac
      841 gggaagttca aagagggcgt tgcctccggc aacctcaaca ctatgttcga atggatgata
      901 ccgacaagca gaggcccaac caaggtaaaa attcatatga agaaagctct gtctggagac
      961 tcttattggg tgtttgtgaa gagagtcggc ggcggctccg gaggaggaga acacgttgcc
     1021 tttggctctg aggatatcga aaatacgctg gccaaaatgg atgacggtca gctggatggc
     1081 ctggcattcg gagccataca gcttgacggc gatggaaaca tcctccagta caacgccgcg
     1141 gagggtgaca ttaccggtcg cgatccaaaa caggttattg gcaagaactt ctttaaggac
     1201 gttgcacccg ggactgactc tcccgaattt tacggaaagt tcaaggaagg ggtggcatct
     1261 ggcaatctta atacaatgtt tgaatggatg attcctacta gtagaggacc aacaaaggtg
     1321 aagatccaca tgaagaaagc cttgtccggc gactcctact gggtctttgt gaaacgggtg
     1381 ggatccgaac aaaagcttat ttctgaagag gacttgtaat aggcggccgc gactctagat
     1441 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1501 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1561 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1621 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     1681 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1741 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     1801 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1861 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     1921 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     1981 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2041 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2101 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2161 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2221 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2281 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2341 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2401 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2461 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2521 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2581 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     2641 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2701 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2761 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2821 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2881 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2941 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3001 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3061 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3121 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3181 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3241 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3301 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3361 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3421 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3481 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3541 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3601 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     3661 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     3721 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3781 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3841 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     3901 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3961 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4201 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4381 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4441 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4501 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4561 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4621 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4681 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4741 tattaccgcc atgcattagt tatt
//
