LOCUS       sequence-396433-        4723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4723)
  AUTHORS   Subach FV, Subach OM, Gundorov IS, Morozova KS, Piatkevich KD, 
            Cuervo AM, Verkhusha VV
  TITLE     Monomeric fluorescent timers that change color from blue to red 
            report on cellular trafficking.
  JOURNAL   Nat Chem Biol. 2009 Feb;5(2):118-26. Epub 2009 Jan 11.
  PUBMED    19136976
REFERENCE   2  (bases 1 to 4723)
  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: "Nat Chem 
            Biol."; date: "2009-02"; volume: "5(2)"; pages: "118-26. Epub 2009 
            Jan 11"
COMMENT     Sequence Label: pSlow-FT-N1
FEATURES             Location/Qualifiers
     source          1..4723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..245
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     202..222
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(247..351)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      378..833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(465..484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     675..696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(851..870)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     905..924
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1083..1793)
                     /codon_start=1
                     /product="blue-to-red fluorescent timer derivative of 
                     mCherry (Subach et al., 2009)"
                     /label=Slow-FT
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHVEGSVNGHEFEIVGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSRAYVKHPADIPDYWKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDVEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1361..1380)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1630..1648
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1801..1881
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(1904..2107)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1933..1953)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2108..2411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2586..3174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2666..2685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3503..3550
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3575..3594
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3782..4576)
                     /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(3782..4576)
                     /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(3893..3912)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4503..4522
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4611..4723,1..245))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4625..4723,1..37)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
       61 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      121 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      181 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
      241 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
      301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
      361 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
      721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcctt
      841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      961 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1021 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
     1081 gcttacttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac
     1141 tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg
     1201 taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcaacgtcg
     1261 tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc
     1321 tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg
     1381 ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc
     1441 agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc
     1501 cacttgaagc cctcggggaa ggacagcttc cagtagtcgg ggatgtcggc ggggtgcttc
     1561 acgtaggccc tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg
     1621 gggccaccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg
     1681 ccctcgccca cgatctcgaa ctcgtggccg ttcacggagc cctccacgtg caccttgaaa
     1741 cgcatgaact ccttgatgat ggccatgtta tcctcctcgc ccttgctcac catggtggcg
     1801 accggtggat cccgggcccg cggtaccgtc gactgcagaa ttcgaagctt gagctcgaga
     1861 tctgagtccg gtagcgctag cggatctgac ggttcactaa accagctctg cttatataga
     1921 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     1981 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     2041 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     2101 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     2161 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     2221 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     2281 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     2341 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     2401 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     2461 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3301 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     3361 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     3421 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     3481 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     3541 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     3601 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     3661 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     3721 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     3781 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     3841 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     3901 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     3961 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     4021 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     4081 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4141 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     4201 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     4261 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     4321 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     4381 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     4441 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     4501 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     4561 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag
     4621 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     4681 tcggcctctg cataaataaa aaaaattagt cagccatggg gcg
//
