LOCUS       sequence-36033-e        3836 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 3836)
  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 3836)
  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: pTRE-Slow-FT
FEATURES             Location/Qualifiers
     source          1..3836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(15..33,34..56,57..75,76..98,99..117,118..140,141..159,
                     160..182,183..201,202..224,225..243,244..266,267..285)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        318..355
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     352..376
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             455..1165
                     /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(600..618)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     868..887
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1173..1307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1197..1216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1251..1270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1733..1752)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1846..1863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2017..2605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2776..3567,3568..3636))
                     /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     3399..3418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3637..3741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3809..3827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      361 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      421 ccgggaccga tccagcctcc gcggccccga attcatggtg agcaagggcg aggaggataa
      481 catggccatc atcaaggagt tcatgcgttt caaggtgcac gtggagggct ccgtgaacgg
      541 ccacgagttc gagatcgtgg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      601 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
      661 gttcatgtac ggctccaggg cctacgtgaa gcaccccgcc gacatccccg actactggaa
      721 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
      781 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
      841 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
      901 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
      961 gctgaagctg aaggacggcg gccactacga cgttgaggtc aagaccacct acaaggccaa
     1021 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     1081 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     1141 cggcatggac gagctgtaca agtaatctag aggatccaga catgataaga tacattgatg
     1201 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1261 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1321 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1381 acctctacaa atgtggtatg gctgattatg atcctgcaag cctcgtcgtc tggccggacc
     1441 acgctatctg tgcaaggtcc ccggacgcgc gctccatgag cagagcgccc gccgccgagg
     1501 caagactcgg gcggcgccct gcccgtccca ccaggtcaac aggcggtaac cggcctcttc
     1561 atcgggaatg cgcgcgacct tcagcatcgc cggcatgtcc cctggcggac gggaagtatc
     1621 agctcgacca agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1681 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1741 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctgcattaat gaatcggcca
     1801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     2221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3781 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
