LOCUS       sequence-61826-e        7357 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 7357)
  AUTHORS   Piatkevich KD, Malashkevich VN, Morozova KS, Nemkovich NA, Almo SC, 
            Verkhusha VV
  TITLE     Extended Stokes shift in fluorescent proteins: chromophore-protein 
            interactions in a near-infrared TagRFP675 variant.
  JOURNAL   Sci Rep. 2013;3:1847. doi: 10.1038/srep01847.
  PUBMED    23677204
REFERENCE   2  (bases 1 to 7357)
  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: "Sci Rep."; 
            date: "2013"; volume: "3:1847. doi"; pages: " 10.1038/srep01847"
COMMENT     Sequence Label: pActinin-TagRFP675-N1
FEATURES             Location/Qualifiers
     source          1..7357
                     /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"
     CDS             3370..4071
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMMYPADGGLEGRNYMALK
                     LVGGGHLICSLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    4194..4315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4231..4250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4285..4304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4322..4777)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4459..4480)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4671..4690
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4804..4908
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4910..5267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4910..5101
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4933..4953)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5118..5253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5302..6096
                     /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             5302..6096
                     /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(5356..5375)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5966..5985
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6284..6303)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6328..6375
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6704..7292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7193..7212
                     /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 caagcttcgc accatcatgg accattatga ttctcagcaa
      661 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      721 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      781 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      841 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      901 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      961 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
     1021 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1081 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1141 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1201 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1261 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1321 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1381 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1441 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1501 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1561 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1621 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1681 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1741 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1801 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1861 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1921 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1981 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     2041 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2101 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2161 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2221 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2281 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2341 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2401 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2461 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2521 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2581 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2641 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2701 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2761 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2821 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2881 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2941 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     3001 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3061 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3121 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3181 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3241 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3301 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3361 gtcgccacca tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
     3421 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     3481 acccagaccc aaagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     3541 ctggctacca gcttcatgta cggcagcaag accttcatca accacaccca gggcatcccc
     3601 gacttctgga agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     3661 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     3721 aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
     3781 ctcggctggg aggccaacac cgagatgatg taccccgctg acggcggcct ggaaggcaga
     3841 aactacatgg ccctgaagct cgtgggcggg ggccacctga tctgcagctt gaagaccaca
     3901 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccgc
     3961 agactggaaa gaatcaagga ggccgacaaa gagacctatg tcgagcagca cgaggtggct
     4021 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta aagcggccgc
     4081 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4141 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     4321 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4381 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4441 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4501 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4561 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4621 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4681 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4741 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4801 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4861 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4921 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4981 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5041 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5101 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5161 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5221 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     5281 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5341 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5401 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5461 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5521 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5581 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5641 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5701 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5761 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5821 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5881 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5941 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6001 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6061 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6121 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6181 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6241 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     6301 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     6361 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     6421 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     6481 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     6541 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6601 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6661 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6781 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6841 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6901 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6961 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7021 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7081 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7141 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7201 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7261 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7321 tcctggcctt ttgctggcct tttgctcaca tgttctt
//
