LOCUS       sequence-244285-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  redox-sensitive red fluorescent protein rxRFP1.4, endoplasmic 
            reticulum localized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   Fan Y, Ai HW
  TITLE     Development of redox-sensitive red fluorescent proteins for imaging 
            redox dynamics in cellular compartments.
  JOURNAL   Anal Bioanal Chem. 2016 Jan 12.
  PUBMED    26758595
REFERENCE   2  (bases 1 to 6338)
  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: "Anal 
            Bioanal Chem. 2016 Jan 12."
COMMENT     Sequence Label: pER-rxRFP1.4
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             721..738
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             742..774
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             787..801
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     1545..1564
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(1657..1675)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1658..1675)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1695..1712)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1701..1925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1971..2399
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2058..2077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2268..2289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2408..2428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2413..2742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2593..2728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2655..2674
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2809..3603
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2863..2882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3473..3492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3777..3898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3814..3833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3868..3887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3947..3963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3960..3982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3971..3987
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3995..4001,4002..4019,4020..4025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4040..4061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4178..4195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4349..4937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4429..4448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5108..5899,5900..5968))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5731..5750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5969..6073)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6148..6167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 atgctgctat ccgtgcccct gctgctcggc ctgctgggcc tggccgccgc cgacggatcc
      721 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
      781 ccgtacgacg atgacgataa ggatccgagc tcgagctctc ctcgttgcgc tataggtcgg
      841 ctgagctcac ccgtggtttc cgagcggatg taccccgagg acggcgccct gaagagcgag
      901 atcaagaagg ggctgaggct gaaggacggc ggccactacg ccgccgaggt caagaccacc
      961 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat caagttggac
     1021 atcgtgtccc acaacgagga ctacaccatc gtggaacagt gcgaacgcgc cgagggccgc
     1081 cactccaccg gcggcatgga cgagctgtac aagggaggta caggcgggag tctggtgagc
     1141 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1201 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1261 gaggcctttc agaccgctaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     1321 gacatcctgt cccctcagtt catgtacggc tccaaggcct acattaagca cccagccgac
     1381 atccccgact acttcaagct gtccttcccc gagggcttca ggtgggagcg cgtgatgaac
     1441 ttcgaggacg gcggcattat tcacgttaac caggactcct ccctgcagga cggcgtattc
     1501 atctacaagg tgaagctgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1561 aagaccatgg gctgggaggc tacgcgtgac caaaggtgca gtcgtaagct tggaggttct
     1621 ggaggaaagg acgagctgta atctagaggg ccctattcta tagtgtcacc taaatgctag
     1681 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1741 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1801 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1861 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1921 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2341 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2401 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2461 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2581 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2641 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2701 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2761 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2821 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2881 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2941 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3001 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3061 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3121 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3181 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3241 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3301 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3361 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3421 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3481 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3541 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3601 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3661 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3721 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3781 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3841 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3901 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3961 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4021 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4081 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4141 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4201 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4261 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5161 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5221 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5281 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5341 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5401 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5461 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5521 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5581 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5641 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5701 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5761 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5821 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5881 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6001 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6061 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6121 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6181 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6241 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6301 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
