LOCUS       sequence-63505-e        5006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5006)
  AUTHORS   Shcherbakova DM, Verkhusha VV
  TITLE     Near-infrared fluorescent proteins for multicolor in vivo imaging.
  JOURNAL   Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521.
  PUBMED    23770755
REFERENCE   2  (bases 1 to 5006)
  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 
            Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521."
COMMENT     Sequence Label: pNLS-iRFP720
FEATURES             Location/Qualifiers
     source          1..5006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      716..725
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             722..1672
                     /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    1794..1915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1831..1850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1885..1904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1922..2377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2059..2080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2271..2290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2404..2508
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2510..2867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2510..2701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2533..2553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2718..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2902..3696
                     /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             2902..3696
                     /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(2956..2975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3566..3585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3884..3903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3928..3975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4304..4892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4793..4812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atgtcgcgga ggcggcacag
      661 ctacgagaac gatggtggac aacctcacaa aaggaggaag acgtctccac cggtcgccac
      721 catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg atgagccgat
      781 ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg ccgacatgac
      841 gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg gcgccctgat
      901 cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga cgatcgcctt
      961 ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga tgcgaaagga
     1021 cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg agctcgagcc
     1081 tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca gcgccatccg
     1141 ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg
     1201 gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact tcagcgggtc
     1261 cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc
     1321 ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac ggatcattcc
     1381 cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca ccgggcggcc
     1441 gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg agttcatgcg
     1501 caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg
     1561 attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg
     1621 cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt aagcggccgc
     1681 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1741 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1801 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1861 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     1921 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1981 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2041 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2101 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2161 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2221 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2281 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2341 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2521 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     2881 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2941 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3001 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3061 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3121 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3181 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3241 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3301 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3361 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3421 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3481 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3541 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3601 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3661 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3721 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3841 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     3901 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     3961 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4021 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4081 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4141 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4201 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4261 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4321 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4381 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4441 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4501 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4561 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4621 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4681 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4741 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4801 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4861 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4921 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4981 tggataaccg tattaccgcc atgcat
//
