LOCUS       sequence-208011-        4941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP in bacterial 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 4941)
  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 Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pBAD-HisD-miRFP
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..290
                     /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     213..232
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             333..350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1347..1364)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1517..1603
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1695..1722
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1741..1832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1833..1901,1902..2693)
                     /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(2051..2070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2864..3452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3353..3372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3606..3623
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3638..3778
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3704..3723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3864..3886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4042..4920)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
       61 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
      121 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
      181 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
      241 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
      301 tgggctaaca ggaggaatta accatgggga gccaccatca ccatcaccat ggcagatcta
      361 tggtcgcagg acacgcaagt ggaagccccg actttggaac cgccgatccg agcgactgtg
      421 agagagaaga aatccatctg gccggatcca tccagccaca cggaacactg ctggtggtct
      481 ccgagcctga ccatagaatc attcaggcat ctgccaacgc cgctgaattc ctgaatctgg
      541 gaagtgtgct gggagtcccc ctggctgaga tcgacgggga tctgctgatc aagattctgc
      601 cccacctgga ccctaccgca gaaggaatgc cagtggccgt caggtgccgc attggaaacc
      661 cctccacaga gtacgatgga ctgatgcatc gaccacctga aggaggactg atcattgagc
      721 tggaacgggc tgggccaccc attgatctga gcggaacact ggctcccgca ctggagcgaa
      781 ttcgaactgc aggatccctg agggctctgt gcgacgatac tgcactgctg ttccagcagt
      841 gtaccggcta cgaccgagtg atggtctatc ggttcgatga acaggggcac ggagaggtgt
      901 attcagaaat acatgtgact ggactggaga gctactttgg caacaggtat cctagctccc
      961 tcgtgccaca gatggcccgg agactgtacg aaagacagag ggtgcgcgtc ctggtggatg
     1021 tctcttatca gcctgtgcca ctggagccaa gactgagtcc cctgactggc agggacctgg
     1081 atatgtcagg gtgttttctg cgctctatga gtcctactca cctgcagttc ctgaagaata
     1141 tgggcgtgcg ggcaacactg gtggtcagcc tggtggtcgg aggcaaactg tgggggctgg
     1201 tgatctgcca ccattatctg ccacgcttca tccattttga gctgcgagca atttgtgagc
     1261 tgctggccga agctatcgca acccgcatta cagccctgta agaattcgaa gcttggctgt
     1321 tttggtggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1381 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1441 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1501 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1561 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1621 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1681 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2041 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3001 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3481 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3541 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3601 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3661 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3721 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3781 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3841 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3901 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     3961 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4021 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4081 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4141 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4201 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4261 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4321 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4381 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4441 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4501 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     4561 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4621 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     4681 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     4741 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     4801 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     4861 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     4921 acttttcata ctcccgccat t
//
