LOCUS       sequence-60518-e        4701 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 4701)
  AUTHORS   Piatkevich KD, Malashkevich VN, Morozova KS, Nemkovich NA, Almo SC, 
            Verkhusha VV
  TITLE     Extended Stokes shift in fluorescent proteins: chromophore-protein 
            interactions in a near-infrared TagRFP675 variant.
  JOURNAL   Sci Rep. 2013;3:1847. doi: 10.1038/srep01847.
  PUBMED    23677204
REFERENCE   2  (bases 1 to 4701)
  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: "Sci Rep."; 
            date: "2013"; volume: "3:1847. doi"; pages: " 10.1038/srep01847"
COMMENT     Sequence Label: pBAD/HisD-TagRFP675
FEATURES             Location/Qualifiers
     source          1..4701
                     /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             328..345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             355..1056
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMMYPADGGLEGRNYMALK
                     LVGGGHLICSLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1102..1119)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1272..1358
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1450..1477
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1496..1587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1588..1656,1657..2448)
                     /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(1806..1825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2619..3207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3108..3127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3361..3378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3393..3533
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3459..3478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3619..3641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3797..4675)
                     /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 ggggagccac catcaccatc accatggcag atctatgagc
      361 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
      421 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gacccaaaga
      481 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
      541 atgtacggca gcaagacctt catcaaccac acccagggca tccccgactt ctggaagcag
      601 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
      661 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
      721 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
      781 aacaccgaga tgatgtaccc cgctgacggc ggcctggaag gcagaaacta catggccctg
      841 aagctcgtgg gcgggggcca cctgatctgc agcttgaaga ccacatacag atccaagaaa
      901 cccgctaaga acctcaagat gcccggcgtc tactatgtgg accgcagact ggaaagaatc
      961 aaggaggccg acaaagagac ctatgtcgag cagcacgagg tggctgtggc cagatactgc
     1021 gacctcccta gcaaactggg gcacaagctt aattaagaat tcgaagcttg gctgttttgg
     1081 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1141 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1201 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1261 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1321 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     1381 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1441 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1501 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1561 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1621 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1681 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1741 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1801 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     1861 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1921 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1981 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2041 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2101 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2161 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2221 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2281 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2341 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2401 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2461 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2521 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2761 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3361 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3421 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3481 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3541 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3601 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3661 aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct
     3721 gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg
     3781 tctgattcgt taccaattat gacaacttga cggctacatc attcactttt tcttcacaac
     3841 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     3901 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     3961 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     4021 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     4081 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     4141 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     4201 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     4261 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     4321 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     4381 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     4441 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     4501 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     4561 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     4621 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     4681 tcatactccc gccattcaga g
//
