LOCUS       sequence-111290-        5603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Marking retinal neurons with infrared fluorescent proteins (iRFP) to
            preserve photoresponsiveness.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  AUTHORS   Fyk-Kolodziej B, Hellmer CB, Ichinose T
  TITLE     Marking cells with infrared fluorescent proteins to preserve 
            photoresponsiveness in the retina.
  JOURNAL   Biotechniques. 2014 Nov 1;57(5):245-53. doi: 10.2144/000114228. 
            eCollection 2014 Nov.
  PUBMED    25391913
REFERENCE   2  (bases 1 to 5603)
  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"; doi: "10.2144/000114228";
            journalName: "Biotechniques"; date: "2014-11-1- 1"; volume: "57"; 
            issue: "5"; pages: "245-53"
COMMENT     Sequence Label: pAAV-CMV-iRFP
FEATURES             Location/Qualifiers
     source          1..5603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        514..716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     667..687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     713..737
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          865..1243
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     regulatory      1335..1344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1341..2291
                     /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="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    2350..2826
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2549..2568)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2866..3006
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2866..2995
                     /label=AAV2 ITR
     rep_origin      3081..3536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3168..3187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3378..3399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3553..3572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3672..3694
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3732..3750)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3818..3922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3923..3991,3992..4783)
                     /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     complement(4141..4160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4954..5542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5443..5462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggagct agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgtcaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      541 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      601 gggagtttgt tttgcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      721 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      781 accgggaccg atccagcctc cgcggattcg aatcccggcc gggaacggtg cattggaacg
      841 cggattcccc gtgccaagag tgacgtaagt accgcctata gagtctatag gcccacaaaa
      901 aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc
      961 tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa
     1021 taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc
     1081 atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta
     1141 ccattctgct tttattttat ggttgggata aggctggatt attctgagtc caagctaggc
     1201 ccttttgcta atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct
     1261 ggtctgtgtg ctggcccatc actttggcaa agaattggga ttcgaacatc gattgaattc
     1321 cccggggatc tgccgccacc atggcggaag gatccgtcgc caggcagcct gacctcttga
     1381 cctgcgacga tgagccgatc catatccccg gtgccatcca accgcatgga ctgctgctcg
     1441 ccctcgccgc cgacatgacg atcgttgccg gcagcgacaa ccttcccgaa ctcaccggac
     1501 tggcgatcgg cgccctgatc ggccgctctg cggccgatgt cttcgactcg gagacgcaca
     1561 accgtctgac gatcgccttg gccgagcccg gggcggccgt cggagcaccg atcactgtcg
     1621 gcttcacgat gcgaaaggac gcaggcttca tcggctcctg gcatcgccat gatcagctca
     1681 tcttcctcga gctcgagcct ccccagcggg acgtcgccga gccgcaggcg ttcttccgcc
     1741 gcaccaacag cgccatccgc cgcctgcagg ccgccgaaac cttggaaagc gcctgcgccg
     1801 ccgcggcgca agaggtgcgg aagattaccg gcttcgatcg ggtgatgatc tatcgcttcg
     1861 cctccgactt cagcggcgaa gtgatcgcag aggatcggtg cgccgaggtc gagtcaaaac
     1921 taggcctgca ctatcctgcc tcaaccgtgc cggcgcaggc ccgtcggctc tataccatca
     1981 acccggtacg gatcattccc gatatcaatt atcggccggt gccggtcacc ccagacctca
     2041 atccggtcac cgggcggccg attgatctta gcttcgccat cctgcgcagc gtctcgcccg
     2101 tccatctgga attcatgcgc aacataggca tgcacggcac gatgtcgatc tcgattttgc
     2161 gcggcgagcg actgtgggga ttgatcgttt gccatcaccg aacgccgtac tacgtcgatc
     2221 tcgatggccg ccaagcctgc gagctagtcg cccaggttct ggcctggcag atcggcgtga
     2281 tggaagagtg agtcgacgcg gccgctcgag cctaagcttg cctcgagcag cgctgctcga
     2341 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2401 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2461 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2521 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2581 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2641 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2701 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     2761 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     2821 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2881 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2941 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3001 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3061 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3121 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3181 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3241 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3301 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3361 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3421 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3481 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3601 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3661 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3721 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3781 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4141 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5101 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5581 ttgctggcct tttgctcaca tgt
//
