LOCUS       sequence-409851-        4734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein mCardinal in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4734)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 4734)
  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 
            Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pBAD-HisD-mCardinal
FEATURES             Location/Qualifiers
     source          1..4734
                     /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..1089
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(1135..1152)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1305..1391
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1483..1510
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1529..1620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1621..1689,1690..2481)
                     /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(1839..1858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2652..3240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3141..3160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3394..3411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3426..3566
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3492..3511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3652..3674
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3830..4708)
                     /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 atctatggtg
      361 agcaagggcg aggagctgat caaggagaac atgcacatga agctgtacat ggaaggcacc
      421 gtgaacaacc accacttcaa gtgcaccacc gaaggggagg gcaagcccta cgagggcacc
      481 cagacccaga ggattaaggt ggtggaggga ggccccctgc cgttcgcatt cgacatcctg
      541 gccacctgct ttatgtacgg gagcaagacc ttcatcaacc acacccaggg catccccgat
      601 ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
      661 gggggcgtgc ttaccgttac ccaggacacc agcctccagg acggctgctt gatctacaac
      721 gtcaagctca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
      781 ggctgggagg ccaccaccga gaccctgtac cccgctgacg gcggcctgga aggcagatgc
      841 gacatggccc tgaagctcgt gggcgggggc cacctgcact gcaacctgaa gaccacatac
      901 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactttgt ggaccgcaga
      961 ctggaaagaa tcaaggaggc cgacaatgag acctacgtcg agcagcacga ggtggctgtg
     1021 gccagatact gcgacctccc tagcaaactg gggcacaaac ttaatggcat ggacgagctg
     1081 tacaagtaag aattcgaagc ttggctgttt tggtggatga gagaagattt tcagcctgat
     1141 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     1201 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1261 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1321 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1381 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1441 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1501 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     1561 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1621 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1681 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1741 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1801 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1861 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1921 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1981 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2041 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2101 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2161 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2221 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2281 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2341 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2401 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2461 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2521 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2581 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2641 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2701 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2761 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3241 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3301 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3361 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3421 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3481 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3541 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3601 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3661 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     3721 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     3781 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     3841 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     3901 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     3961 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     4021 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     4081 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     4141 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     4201 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     4261 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     4321 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     4381 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     4441 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     4501 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     4561 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     4621 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     4681 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agag
//
