LOCUS       sequence-60544-e        6379 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 6379)
  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 6379)
  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: pPaxillin-TagRFP675-N1
FEATURES             Location/Qualifiers
     source          1..6379
                     /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"
     misc_feature    2277..2329
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2343..3044
                     /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    3167..3288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3204..3223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3258..3277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3295..3750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3432..3453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3644..3663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3777..3881
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3883..4240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3883..4074
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3906..3926)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4091..4226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4275..5069
                     /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             4275..5069
                     /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(4329..4348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4939..4958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5257..5276)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5301..5348
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5677..6265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6166..6185
                     /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 gctagcacca
      601 tggacgacct cgatgcctta ctggcagacc tggaatccac cacctcgcat atctctaagc
      661 gaccggtgtt tctaacagag gaaacgcctt actcctaccc aactggaaac catacgtacc
      721 aggagattgc tgtgccaccg ccggtgcccc cgccgccttc cagtgaggcc ctaaatggca
      781 ctgtgattga ccccttagac cagtggcaac ccagcgtgtc cagatacggc caccagcaac
      841 ctcagtccca gtctccgata tacagctcta gtgccaaaag ctccagtgcc tctgttccta
      901 gagacgggct cagctcccct tctccccgtg ccagtgagga ggaacacgtg tacagcttcc
      961 caaacaagca gaagtctgca gaaccatctc ccacaatgac cagcacctcc ctgggcagca
     1021 acctctcgga actggacagg cttctcctgg aactgaatgc tgttcaacat aatcccccca
     1081 gtggcttctc agcagatgaa gtcagcagaa gcccatcact gcccaatgtg accggacctc
     1141 actatgtcat cccagagagc agcagctctg caggagggaa ggctgcaccc cctacaaaag
     1201 aaaaaccaaa gcggaatggt gggcgtggga ttgaagatgt gcgtcccagt gtggagagcc
     1261 tgctggatga gctggagagc tctgtgccaa gcccagtccc tgcaatcact gtgagccaag
     1321 gggaggtgag cagcccccag cgtgtcaacg ccagtcagca gcagacccgt atctctgctt
     1381 cttcagctac acgagaactg gatgagctga tggcgtccct ctctgacttt aagttcatgg
     1441 cacaggggaa agccggcggc agcagctccc caccatccac caccccaaag cctggcagtc
     1501 agctggacac catgctggga agtctgcagt ctgacctgaa caaactgggg gtagcgacgg
     1561 tcgccaaggg tgtgtgcgga gcctgcaaga agccgattgc tgggcaggta gttacagcca
     1621 tggggaagac ctggcaccct gagcacttcg tctgcaccca ctgccaggag gagattggat
     1681 cacggaactt ctttgagcgg gatgggcagc cctactgtga gaaggactat cacaacctct
     1741 tctctcctcg ctgctactac tgcaatgggc caatccttga taaagtggtg acggctttgg
     1801 acaggacgtg gcaccctgaa cactttttct gtgcccagtg tggagttttc tttggacctg
     1861 aaggatttca tgagaaggat ggcaaagcct attgccgcaa ggactacttt gacatgtttg
     1921 ctcccaagtg tggaggctgt gcccgggcta tcctggaaaa ttacatttct gccttaaaca
     1981 ccctgtggca ccctgagtgc tttgtctgtc gggaatgttt cacaccgttc atcaacggca
     2041 gcttcttcga gcacgacggc cagccctact gcgaggtgca ttaccacgag cgccgcggct
     2101 cgctgtgctc tggctgccag aagcccatca cgggacgctg catcactgcc atgggcaaga
     2161 aatttcaccc cgaacacttt gtctgtgcct tctgcctcaa gcagctcaac aaaggaacct
     2221 tcaaagaaca gaacgacaag ccctactgtc agaactgctt tctcaaactc ttctgtctcg
     2281 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2341 ccatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga
     2401 acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     2461 cccaaagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     2521 ccagcttcat gtacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct
     2581 ggaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
     2641 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
     2701 agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct
     2761 gggaggccaa caccgagatg atgtaccccg ctgacggcgg cctggaaggc agaaactaca
     2821 tggccctgaa gctcgtgggc gggggccacc tgatctgcag cttgaagacc acatacagat
     2881 ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac cgcagactgg
     2941 aaagaatcaa ggaggccgac aaagagacct atgtcgagca gcacgaggtg gctgtggcca
     3001 gatactgcga cctccctagc aaactggggc acaagcttaa ttaaagcggc cgcgactcta
     3061 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3121 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3301 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3361 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3421 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3481 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3541 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3601 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3661 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3721 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3781 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3841 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3901 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3961 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4021 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4081 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4141 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4201 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4261 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4321 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4381 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4441 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4501 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4561 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4621 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4681 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4741 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4801 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4861 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4921 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4981 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5041 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5101 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5161 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5221 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5281 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5341 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5401 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5461 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6361 ccgtattacc gccatgcat
//
