LOCUS       sequence-478804-        5529 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Expresses P450-td8ox2StayGold in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 5529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: pCytERM-td8ox2StayGold-N1
FEATURES             Location/Qualifiers
     source          1..5529
                     /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      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2317..2438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2354..2373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2408..2427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2445..2900)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2582..2603)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2794..2813
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2927..3031
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3033..3390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3033..3224
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3056..3076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3241..3376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3425..4219
                     /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             3425..4219
                     /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(3479..3498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4089..4108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4407..4426)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4451..4498
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4827..5415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5316..5335
                     /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 atctatggac cctgtggtgg tgctggggct ctgtctctcc tgtttgcttc
      661 tcctttcact ctggaaacag agctatgggg gagggaagct tcgaattctg cagtcgacgg
      721 taccgcgggc ccgggatcca ccggtcgcca ccatggctag cactggggaa gaactgttca
      781 caggagtcgt gccgtttaag ttccagttga agggcacgat caatggaaag tctttcacag
      841 tggaaggaga gggtgagggt aatagccatg aaggttcaca caaaggcaag tacgtctgta
      901 cctctggaaa gcttccgatg tcatgggccg ctctgggtac ctcctttggg tatggcatga
      961 aatactatac aaagtacccc tcagggttga agaactggtt ccatgaggtc atgcccgaag
     1021 gattcaccta tgatcgccac atccagtata agggcgacgg ctctattcat gccaaacatc
     1081 aacacttcat gaagaacggc acctaccata acatcgtgga gtttactgga caagacttta
     1141 aggagaatag cccagttctg accggtgata tgaacgtaag cctgcctaac gaagtgcagc
     1201 acattcccag ggacgatggg gttgagtgtc ctgttaccct gctgtaccct ctgttgagcg
     1261 acaaatccaa atgcgtagag gcgtttcaga acaccattat caaaccactg cataaccaac
     1321 cagctcccga tgtaccgtac cattggatac ggaaacagta tacccagtca aaagacgaca
     1381 ctgaggaacg tgaccacatt atccagtccg agacacttga agcacacctc tattcacgaa
     1441 caaaactgga ggggcatggc actggctcta ctggatccgg ctccagtgga acagcatcaa
     1501 gcgaggacaa caatatggcc tccacaggag aggagctgtt caccggagtc gttccattca
     1561 aatttcagct gaaaggcacc attaatggga aaagcttcac agtggaggga gaaggtgaag
     1621 gtaatagtca cgaagggagc cacaagggaa aatacgtctg cactagcggg aaactcccaa
     1681 tgtcttgggc tgcactgggg actagctttg gctacgggat gaagtactac acgaagtatc
     1741 cttccggcct caagaactgg tttcacgagg tgatgccaga agggtttacc tatgatagac
     1801 acatacagta caagggcgac ggcagtatcc acgcgaaaca ccagcatttc atgaagaatg
     1861 ggacgtacca caacattgtg gaatttacgg gccaagactt caaggagaat tctcccgtgc
     1921 ttaccggcga tatgaacgtg agtctgccaa atgaggtgca gcatatacct agggacgatg
     1981 gtgtggaatg tcccgtcaca ctgctgtatc ctctgctcag tgataaatct aagtgcgttg
     2041 aggccttcca aaacaccatc atcaagccct tgcataatca gcctgcccct gatgtgccct
     2101 atcactggat ccggaagcag tacacacagt ccaaggatga cactgaggag agagatcaca
     2161 ttatccagag cgagactctc gaagcccacc tttaagcggc cgcgactcta gatcataatc
     2221 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2281 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2341 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2401 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2461 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2521 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2581 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2641 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2701 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2761 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2821 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2881 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2941 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3001 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3241 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3301 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3361 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3421 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3481 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3541 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3601 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3661 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3721 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3781 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3841 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3901 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3961 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4021 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4081 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4141 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4201 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4261 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4321 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4381 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4441 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4501 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4561 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4621 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4681 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4741 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4801 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4861 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4921 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4981 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5041 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5101 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5161 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5221 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5281 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5341 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5401 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5461 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5521 gccatgcat
//
