LOCUS       sequence-387504-        5703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of miRFP720 and HO1 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5703)
  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 5703)
  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-miRFP720-HO1
FEATURES             Location/Qualifiers
     source          1..5703
                     /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..1305
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     primer_bind     complement(2104..2121)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2274..2360
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2452..2479
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2498..2589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2590..2658,2659..3450)
                     /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(2808..2827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3621..4209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4110..4129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4363..4380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4395..4535
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4461..4480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4621..4643
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4799..5677)
                     /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 accatggcct cgagatggcg
      361 gaaggatccg tcgccaggca gcctgacctc ttgacctgcg acgatgagcc gatccatatc
      421 cccggtgcca tccaaccgca tggactgctg ctcgccctcg ccgccgacat gacgatcgtt
      481 gccggcagcg acaaccttcc cgaactcacc ggactggcga tcggcgccct gatcggccgc
      541 tctgcggccg atgtcttcga ctcggagacg cacaaccgtc tgacgatcgc cttggccgag
      601 cccggggcgg ccgtcggagc accgatcact gtcggcttca cgatgcgaaa ggacgcaggc
      661 ttcatcggct cctggcatcg ccatgatcag ctcatcttcc tcgagctcga gcctccccag
      721 cgggacgtcg ccgagccgca ggcgttcttc cgccgcacca acagcgccat ccgccgcctg
      781 caggccgccg aaaccttgga aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt
      841 accggcttcg atcgggtgat gatctatcgc ttcgcctccg acttcagcgg gtccgtgatc
      901 gcagaggatc ggtgcgccga ggtcgagtca aaactaggcc tgcactatcc tgcctcattc
      961 atcccggcgc aggcccgtcg gctctatacc atcaacccgg tacggatcat tcccgatatc
     1021 aattatcggc cggtgccggt caccccagac ctcaatccgg tcaccgggcg gccgattgat
     1081 cttagcttcg ccatcctgcg cagcgtctcg cccaaccatc tggagttcat gcgcaacata
     1141 ggcatgcacg gcacgatgtc gatctcgatt ttgcgcggcg agcgactgtg gggattgatc
     1201 gtttgccatc accgaacgcc gtactacgtc gatctcgatg gccgccaagc ctgcaagaga
     1261 gtcgccgaga gactggccac ccagatcggc gtgatggaag agtaatagct gcagaagaag
     1321 gagatataca tacccatgag tgtcaactta gcttcccagt tgcgggaagg gacgaaaaaa
     1381 tcccactcca tggcggagaa cgtcggcttt gtcaaatgct tcctcaaggg cgttgtcgag
     1441 aaaaattcct accgtaagct ggttggcaat ctctactttg tctacagtgc catggaagag
     1501 gaaatggcaa aatttaagga ccatcccatc ctcagccaca tttacttccc cgaactcaac
     1561 cgcaaacaaa gcctagagca agacctgcaa ttctattacg gctccaactg gcggcaagaa
     1621 gtgaaaattt ctgccgctgg ccaagcctat gtggaccgag tccggcaagt ggccgctacg
     1681 gcccctgaat tgttggtggc ccattcctac acccgttacc tgggggatct ttccggcggt
     1741 caaattctca agaaaattgc ccaaaatgcc atgaatctcc acgatggtgg cacagctttc
     1801 tatgaatttg ccgacattga tgacgaaaag gcttttaaaa atacctaccg tcaagctatg
     1861 aatgatctgc ccattgacca agccaccgcc gaacggattg tggatgaagc caatgacgcc
     1921 tttgccatga acatgaaaat gttcaacgaa cttgaaggca acctgatcaa ggcgatcggc
     1981 attatggtgt tcaacagcct cacccgtcgc cgcagtcaag gcagcaccga agttggcctc
     2041 gccacctccg aaggctaaga attcgaagct tggctgtttt ggtggatgag agaagatttt
     2101 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     2161 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     2221 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     2281 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2341 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     2401 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     2461 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     2521 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2581 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2641 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2701 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2761 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2821 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2881 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2941 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3001 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3061 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3121 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3181 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3241 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3301 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3361 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3421 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3481 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3541 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4261 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4321 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4381 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4441 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4501 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4561 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4621 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga
     4681 tcaattcgcg cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag
     4741 ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt
     4801 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
     4861 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
     4921 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
     4981 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     5041 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     5101 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     5161 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     5221 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     5281 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     5341 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     5401 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     5461 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     5521 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     5581 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     5641 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca
     5701 gag
//
