LOCUS       sequence-330961-        4568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4568)
  AUTHORS   Kolber NS, Fattal R, Bratulic S, Carver GD, Badran AH
  TITLE     Orthogonal translation enables heterologous ribosome engineering in 
            E. coli.
  JOURNAL   Nat Commun. 2021 Jan 26;12(1):599. doi: 10.1038/s41467-020-20759-z.
  PUBMED    33500394
REFERENCE   2  (bases 1 to 4568)
  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 
            Commun."; date: "2021-01-26"; volume: "12(1):599. doi"; pages: " 
            10.1038/s41467-020-20759-z"
COMMENT     Sequence Label: pNK055b
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..291
                     /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     214..233
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             330..1040
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(475..493)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     743..762
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     937..960
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     terminator      complement(1067..1098)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1114..1773)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(1888..2838)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2886..3108
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3670..4548)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
       61 ggctcttctc gctaacccaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
      121 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
      181 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
      241 tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt
      301 acctgcaggt gcagtaggag gaaaaaaaaa tggtgagcaa gggcgaggag gataacatgg
      361 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
      421 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
      481 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttca
      541 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
      601 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
      661 ccgtgaccca ggactcctcc ctacaggacg gcgagttcat ctacaaggtg aagctgcgcg
      721 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
      781 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
      841 agctgaagga cggcggccac tacgacgccg aggtcaagac cacctacaag gccaagaagc
      901 ccgtgcagct gcccggcgcc tacaacgtca acatcaagct ggacatcacc tcccacaacg
      961 aggactacac catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcggca
     1021 tggacgagct gtacaagtaa acttaattaa cggcactcct cagccaagtc aaaagcctcc
     1081 ggtcggaggc ttttgactac atgcccatgg cgtttacgcc ccgccctgcc actcatcgca
     1141 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     1201 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatagt
     1261 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     1321 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     1381 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     1441 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     1501 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaactc
     1561 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     1621 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     1681 acattgagta actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     1741 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     1801 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     1861 tcttacgtgc caggccaaat aggccgttca gatccttccg tatttagcca gtatgttctc
     1921 tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catgcttact
     1981 ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc
     2041 atcgtgtagt gtttttctta gtccgttacg taggtaggaa tctgatgtaa tggttgttgg
     2101 tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg
     2161 tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac
     2221 caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt
     2281 aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag
     2341 gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc
     2401 ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat
     2461 gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt
     2521 ttcgcttgag aacttggcat agtttgtcca ctggaaaatc ttaaagcctt taaccaaagg
     2581 attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata
     2641 agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt
     2701 ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat
     2761 tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa
     2821 aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt
     2881 gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa
     2941 attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc
     3001 tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa
     3061 agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc
     3121 agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc
     3181 ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc
     3241 ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg
     3301 cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt ttatggattc
     3361 atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca gggcgtttta
     3421 tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg
     3481 attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca
     3541 cccagtaagg cagcggtatc atcaacaggc ttacccgtct tactgtcaag aggacatccg
     3601 gtcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttgct gaggagactt
     3661 agggaccctt tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     3721 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     3781 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     3841 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     3901 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     3961 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     4021 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     4081 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     4141 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     4201 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     4261 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     4321 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     4381 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     4441 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     4501 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     4561 cccaccat
//
