LOCUS       sequence-192527-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   Mezzanotte L, Iljas JD, Que I, Chan A, Kaijzel E, Hoeben R, Lowik C
  TITLE     Optimized longitudinal monitoring of stem cell grafts in mouse brain
            using a novel bioluminescent/near infrared fluorescent fusion 
            reporter.
  JOURNAL   Cell Transplant. 2017 Apr 26. doi: 10.3727/096368917X695407.
  PUBMED    28447574
REFERENCE   2  (bases 1 to 6565)
  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: "Cell 
            Transplant. 2017 Apr 26. doi: 10.3727/096368917X695407."
COMMENT     Sequence Label: pluc2-iRFP720
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      39..627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     528..547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        802..1105
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1106..1309
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1260..1280
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3016..3025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3022..3972
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    4094..4215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4131..4150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4185..4204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4222..4677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4359..4380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4571..4590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4704..4808
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4810..5167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4810..5001
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4833..4853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5018..5153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5202..5996
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5202..5996
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5256..5275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5866..5885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6184..6203)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6228..6275
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
       61 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      721 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
      781 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      841 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      901 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      961 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1021 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1081 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1141 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1201 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1261 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     1321 ccgtcagatc cgctagcaat ggaagatgcc aaaaacatta agaagggccc agcgccattc
     1381 tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc
     1441 ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc
     1501 gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca
     1561 aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt
     1621 gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg
     1681 ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa
     1741 aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc
     1801 aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc
     1861 ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg
     1921 atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc
     1981 gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac
     2041 accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc
     2101 tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg
     2161 cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc
     2221 ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc
     2281 ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca
     2341 ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa
     2401 ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg
     2461 gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc
     2521 cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag
     2581 gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc
     2641 gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg
     2701 gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac
     2761 gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc
     2821 gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt
     2881 ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag
     2941 atccgcgaga ttctcattaa ggccaagaag ggcggcaagc agtcgacggt accgcgggcc
     3001 cgggatccac cggtcgccac catggcggaa ggatccgtcg ccaggcagcc tgacctcttg
     3061 acctgcgacg atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc
     3121 gccctcgccg ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga
     3181 ctggcgatcg gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac
     3241 aaccgtctga cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc
     3301 ggcttcacga tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc
     3361 atcttcctcg agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc
     3421 cgcaccaaca gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc
     3481 gccgcggcgc aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc
     3541 gcctccgact tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa
     3601 ctaggcctgc actatcctgc ctcattcatc ccggcgcagg cccgtcggct ctataccatc
     3661 aacccggtac ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc
     3721 aatccggtca ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc
     3781 aaccatctgg agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg
     3841 cgcggcgagc gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat
     3901 ctcgatggcc gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg
     3961 atggaagagt aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     4021 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4081 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4141 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4201 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4261 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4321 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4381 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4441 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4501 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4561 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4621 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4801 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5101 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5161 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5221 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5281 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5341 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5401 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5461 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5521 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5581 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5641 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5701 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5761 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5821 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5881 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5941 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6001 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6061 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6181 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6241 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6301 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6361 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6421 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6481 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6541 cccttaacgt gagttttcgt tccac
//
