LOCUS       sequence-194618-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid that express membrane-targeted iRFP under the CMV 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  AUTHORS   Richie CT, Whitaker LR, Whitaker KW, Necarsulmer J, Baldwin HA, 
            Zhang Y, Fortuno L, Hinkle JJ, Koivula P, Henderson MJ, Sun W, Wang 
            K, Smith JC, Pickel J, Ji N, Hope BT, Harvey BK
  TITLE     Near-infrared fluorescent protein iRFP713 as a reporter protein for 
            optogenetic vectors, a transgenic Cre-reporter rat, and other 
            neuronal studies.
  JOURNAL   J Neurosci Methods. 2017 Jun 1;284:1-14. doi: 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2.
  PUBMED    28380331
REFERENCE   2  (bases 1 to 6091)
  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: "J Neurosci 
            Methods."; date: "2017-06-1"; volume: "284:1-14. doi"; pages: " 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2"
COMMENT     Sequence Label: pOTTC591	- pCMV Mem-iRFP
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        464..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     618..638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    690..770
                     /label=MCS
                     /note="multiple cloning site"
     CDS             850..1797
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="AEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSD
                     NLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIG
                     SWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITG
                     FDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDIN
                     YRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGLI
                     VCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    1919..2040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2010..2029
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2047..2502)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2184..2205)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2396..2415
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2529..2633
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2635..2992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2635..2826
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2658..2678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2843..2978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3000..4025
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3000..4025
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4142..4223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4251..4269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4337..4441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4442..4510,4511..5302)
                     /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(4660..4679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5488..6076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5977..5996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
       61 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
      121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      661 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc
      721 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
      781 ctgtgctgta tgagaagaac caaacaggtt gaaaagaatg atgaggacca aaagatcagc
      841 aagggcgccg ctgaaggatc cgtcgccagg cagcctgacc tcttgacctg cgacgatgag
      901 ccgatccata tccccggtgc catccaaccg catggactgc tgctcgccct cgccgccgac
      961 atgacgatcg ttgccggcag cgacaacctt cccgaactca ccggactggc gatcggcgcc
     1021 ctgatcggcc gctctgcggc cgatgtcttc gactcggaga cgcacaaccg tctgacgatc
     1081 gccttggccg agcccggggc ggccgtcgga gcaccgatca ctgtcggctt cacgatgcga
     1141 aaggacgcag gcttcatcgg ctcctggcat cgccatgatc agctcatctt cctcgagctc
     1201 gagcctcccc agcgggacgt cgccgagccg caggcgttct tccgccgcac caacagcgcc
     1261 atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct gcgccgccgc ggcgcaagag
     1321 gtgcggaaga ttaccggctt cgatcgggtg atgatctatc gcttcgcctc cgacttcagc
     1381 ggcgaagtga tcgcagagga tcggtgcgcc gaggtcgagt caaaactagg cctgcactat
     1441 cctgcctcaa ccgtgccggc gcaggcccgt cggctctata ccatcaaccc ggtacggatc
     1501 attcccgata tcaattatcg gccggtgccg gtcaccccag acctcaatcc ggtcaccggg
     1561 cggccgattg atcttagctt cgccatcctg cgcagcgtct cgcccgtcca tctggaattc
     1621 atgcgcaaca taggcatgca cggcacgatg tcgatctcga ttttgcgcgg cgagcgactg
     1681 tggggattga tcgtttgcca tcaccgaacg ccgtactacg tcgatctcga tggccgccaa
     1741 gcctgcgagc tagtcgccca ggttctggcc tggcagatcg gcgtgatgga agagtgagcg
     1801 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1861 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1921 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1981 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2041 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2101 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2161 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2221 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2281 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2341 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2401 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2461 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2521 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2581 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2641 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2761 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2821 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2881 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2941 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgata
     3001 tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     3061 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     3121 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     3181 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     3241 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     3301 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     3361 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     3421 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     3481 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     3541 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     3601 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     3661 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     3721 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     3781 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     3841 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     3901 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     3961 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     4021 aatagagttc tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4081 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4141 taacttgttt attgcagctt gtaatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcctcgcga tccacaacca tggcgaaagg gcctcgtgat
     4261 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4321 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4381 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4441 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4501 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4561 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4621 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4681 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4741 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4801 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4861 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4921 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4981 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5041 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5101 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5161 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5221 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5281 ctcactgatt aagcattggt aaggcgcgcc aggccctgcc atagcctcag gttactcata
     5341 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5401 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5881 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6061 gcggagccta tggaaaaacg ccagcaacgc g
//
