LOCUS       sequence-36035-e        4785 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4785)
  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 4785)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pBAD/HisB-Medium-FT
FEATURES             Location/Qualifiers
     source          1..4785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             430..1140
                     /codon_start=1
                     /product="blue-to-red fluorescent timer derivative of 
                     mCherry (Subach et al., 2009)"
                     /label=Medium-FT
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHLEGSVDGHEFEIEGEGEGRPYEG
                     TQSAKLKVTKGGPLPFAWDILSPQFMYGSRAYVKHPADIPDYWKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERIYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVLLPGAYNVNIKMDITSHNEDYTIVEQCERA
                     EGHHSTGGMDELYK"
     primer_bind     complement(575..593)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     843..862
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1186..1203)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1356..1442
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1534..1561
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1580..1671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1672..1740,1741..2532)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1890..1909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3445..3462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3477..3617
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3543..3562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3703..3725
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3881..4759)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagatcta tggtaagcaa gggcgaggag gataacatgg ccatcatcaa ggaattcatg
      481 cgtttcaagg tgcacctgga gggctccgtg gacggccacg agttcgagat cgagggcgag
      541 ggcgagggcc gcccctacga gggcacccag agcgccaagc tgaaggtgac caagggtggc
      601 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc cagggcctac
      661 gtgaagcacc ccgccgacat ccccgactac tggaagctgt ccttccccga gggcttcaag
      721 tgggagcgcg tgatgaactt cgaggatggc ggcgtggtga ccgtgaccca ggactcctcc
      781 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccttccgac
      841 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatatacccc
      901 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
      961 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgctgct gcccggcgcc
     1021 tacaacgtca acatcaagat ggacatcacc tcccacaacg aggactacac catcgttgaa
     1081 cagtgcgaac gcgccgaggg ccaccattcc accggcggca tggacgagct gtacaagtaa
     1141 gaattcgaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1201 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1261 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1321 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1381 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1441 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1501 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1561 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1621 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1681 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1741 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1801 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1861 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     1921 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1981 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2041 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2101 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2161 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2221 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2581 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3421 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     3541 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3601 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3661 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3721 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
     3781 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     3841 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     3901 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     3961 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     4021 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     4081 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     4141 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4201 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     4261 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     4321 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     4381 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     4441 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     4501 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     4561 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     4621 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     4681 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     4741 cattttgcgc ttcagccata cttttcatac tcccgccatt cagag
//
