LOCUS       sequence-313677-        4736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U1A-dependent translational repression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4736)
  AUTHORS   Kawasaki S, Fujita Y, Nagaike T, Tomita K, Saito H
  TITLE     Synthetic mRNA devices that detect endogenous proteins and 
            distinguish mammalian cells.
  JOURNAL   Nucleic Acids Res. 2017 May 19. doi: 10.1093/nar/gkx298.
  PUBMED    28525643
REFERENCE   2  (bases 1 to 4736)
  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: "Nucleic 
            Acids Res. 2017 May 19. doi: 10.1093/nar/gkx298."
COMMENT     Sequence Label: pU1utr-EGFP
FEATURES             Location/Qualifiers
     source          1..4736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(622..624,625..627,628..1341)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(622..624,625..627,628..1338)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(667..688)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(928..947)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1275..1296
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1464..1585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1501..1520)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1555..1574
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1592..2047)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1729..1750)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1941..1960
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2074..2178
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2180..2537
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2180..2371
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2203..2223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2388..2523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2572..3366
                     /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             2572..3366
                     /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(2626..2645)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3236..3255
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3554..3573)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3598..3645
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3974..4562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4463..4482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atccacagca ttgtacccag agtctgtccc cagacattgc acctggcgct
      601 gtagatctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      661 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      721 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      781 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      841 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      901 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      961 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1021 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1081 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1141 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1201 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1261 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1321 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     1381 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1441 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1501 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1561 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     1621 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1681 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1741 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1801 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1861 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1921 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1981 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2041 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2101 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2161 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2461 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2521 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     2581 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2641 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2701 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     2761 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2821 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2881 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2941 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3001 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3061 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3121 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3181 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3241 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3301 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3361 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3421 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3481 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     3541 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3601 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     3661 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     3721 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     3781 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     3841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4081 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4561 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4621 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     4681 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
