LOCUS       sequence-243877-        4974 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 4974)
  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 4974)
  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: pMito-iRFP670
FEATURES             Location/Qualifiers
     source          1..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             619..693
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      721..730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             727..1662
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    1784..1905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1821..1840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1875..1894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1912..2367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2049..2070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2261..2280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2394..2498
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2500..2857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2500..2691
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2523..2543)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2708..2843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2892..3686
                     /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             2892..3686
                     /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(2946..2965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3556..3575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3874..3893)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3918..3965
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4294..4882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4783..4802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcat gtccgtcctg acgccgctgc tgctgcgggg cttgacaggc
      661 tcggcccggc ggctcccagt gccgcgcgcc aagatccatt cgttggggga tccaccggtc
      721 gccaccatgg cgcgtaaggt cgatctcacc tcctgcgatc gcgagccgat ccacatcccc
      781 ggcagcattc agccgtgcgg ctgcctgcta gcctgcgacg cgcaggcggt gcggatcacg
      841 cgcattacgg aaaatgccgg cgcgttcttt ggacgcgaaa ctccgcgggt cggtgagcta
      901 ctcgccgatt acttcggcga gaccgaagcc catgcgctgc gcaacgcact ggcgcagtcc
      961 tccgatccaa agcgaccggc gctgatcttc ggttggcgcg acggcctgac cggccgcacc
     1021 ttcgacatct cactgcatcg ccatgacggt acatcgatca tcgagttcga gcctgcggcg
     1081 gccgaacagg ccgacaatcc gctgcggctg acgcggcaga tcatcgcgcg caccaaagaa
     1141 ctgaagtcgc tcgaagagat ggccgcacgg gtgccgcgct atctgcaggc gatgctcggc
     1201 tatcaccgcg tgatgttgta ccgcttcgcg gacgacggct ccgggatggt gatcggcgag
     1261 gcgaagcgca gcgacctcga gagctttctc ggtcagcact ttccggcgtc gctggtcccg
     1321 cagcaggcgc ggctactgta cttgaagaac gcgatccgcg tggtctcgga ttcgcgcggc
     1381 atcagcagcc ggatcgtgcc cgagcacgac gcctccggcg ccgcgctcga tctgtcgttc
     1441 gcgcacctgc gcagcatctc gccctgccat ctcgaatttc tgcggaacat gggcgtcagc
     1501 gcctcgatgt cgctgtcgat catcattgac ggcacgctat ggggattgat catctgtcat
     1561 cattacgagc cgcgtgccgt gccgatggcg cagcgcgtcg cggccgaaat gttcgccgac
     1621 ttcttatcgc tgcacttcac cgccgcccac caccaacgct aagcggccgc gactctagat
     1681 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1741 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1801 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1861 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     1921 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1981 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2041 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2101 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2161 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2221 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2281 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2341 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2401 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2461 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2521 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2581 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2641 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2701 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2761 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2821 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     2881 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2941 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3001 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3061 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3121 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3181 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3241 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3301 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3361 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3421 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3481 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3541 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3601 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3661 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3721 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3781 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3841 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     3901 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     3961 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4021 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4081 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4141 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4201 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4261 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4321 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4381 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4441 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4501 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4561 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4621 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4681 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4741 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4801 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4861 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4921 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tgga
//
