LOCUS       sequence-204538-        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        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"
     primer_bind     complement(1342..1359)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1512..1598
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1690..1717
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1736..1827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1828..1896,1897..2688)
                     /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(2046..2065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2859..3447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3348..3367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3601..3618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3633..3773
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3699..3718)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3859..3881
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4037..4915)
                     /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 atctatggtc
      361 gcaggacacg caagtggaag ccccgacttt ggaaccgccg atccgagcga ctgtgagaga
      421 gaagaaatcc atctggccgg atccatccag ccacacggaa cactgctggt ggtctccgag
      481 cctgaccata gaatcattca ggcatctgcc aacgccgctg aattcctgaa tctgggaagt
      541 gtgctgggag tccccctggc tgagatcgac ggggatctgc tgatcaagat tctgccccac
      601 ctggacccta ccgcagaagg aatgccagtg gccgtcaggt gccgcattgg aaacccctcc
      661 acagagtacg atggactgat gcatcgacca cctgaaggag gactgatcat tgagctggaa
      721 cgggctgggc cacccattga tctgagcgga acactggctc ccgcactgga gcgaattcga
      781 actgcaggat ccctgagggc tctgtgcgac gatactgcac tgctgttcca gcagtgtacc
      841 ggctacgacc gagtgatggt ctatcggttc gatgaacagg ggcacggaga ggtgtattca
      901 gaaatacatg tgactggact ggagagctac tttggcaaca ggtatcctag ctccctcgtg
      961 ccacagatgg cccggagact gtacgaaaga cagagggtgc gcgtcctggt ggatgtctct
     1021 tatcagcctg tgccactgga gccaagactg agtcccctga ctggcaggga cctggatatg
     1081 tcagggtgtt ttctgcgctc tatgagtcct actcacctgc agttcctgaa gaatatgggc
     1141 gtgcgggcaa cactggtggt cagcctggtg gtcggaggca aactgtgggg gctggtgatc
     1201 tgccaccatt atctgccacg cttcatccat tttgagctgc gagcaatttg tgagctgctg
     1261 gccgaagcta tcgcaacccg cattacagcc ctgtaagaat tcgaagcttg gctgttttgg
     1321 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1381 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1441 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1501 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1561 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     1621 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1681 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1981 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2041 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3001 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3301 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3361 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3421 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3481 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3541 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3601 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3661 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3721 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3781 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3841 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3901 aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct
     3961 gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg
     4021 tctgattcgt taccaattat gacaacttga cggctacatc attcactttt tcttcacaac
     4081 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     4141 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     4201 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     4261 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     4321 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     4381 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     4441 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     4501 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     4561 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     4621 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     4681 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     4741 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     4801 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     4861 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     4921 tcatactccc gccattcaga g
//
