LOCUS       sequence-47078-e        8384 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 8384)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 8384)
  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: "Mol Cell.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pT7-EGFP-C1-HsPAN2
FEATURES             Location/Qualifiers
     source          1..8384
                     /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"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5172..5293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5209..5228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5263..5282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5300..5755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5437..5458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5649..5668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5782..5886
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5888..6245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5888..6079
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5911..5931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6096..6231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6280..7074
                     /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             6280..7074
                     /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(6334..6353)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6944..6963
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7262..7281)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7306..7353
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7682..8270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8171..8190
                     /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 gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctatgaactt tgagggtctg gaccctgaac tggcagaata
     1441 tgccccagcc atgcattctg ccctggaccc tgtcttggat gcccacctga acccaagtct
     1501 gctacagaat gtggagctgg acccagaggg agtggccttg gaggctcttc ccgtccagga
     1561 atcagtgcac ataatggaag gtgtctactc tgaattgcac agcgtggtgg ctgaagtggg
     1621 tgtacctgtt tccgtctccc actttgactt gcacgaggag atgctgtggg tggggagcca
     1681 cgggggccat gccacttcat tttttggccc agccttggag cgctactcat cctttcaagt
     1741 caatggcagt gatgatattc ggcagatcca gagcctggag aatggtatcc tttttctcac
     1801 caagaacaac ctcaagtata tggcccgtgg gggcctcatt atatttgatt acctgctgga
     1861 tgagaatgag gatatgcaca gtctcctact gactgacagc agcactctac tcgttggtgg
     1921 gctgcagaat cacatactag agattgatct taacactgtc caggagactc agaagtatgc
     1981 agtagagacg cctggagtca ccatcatgag acagacaaat cgcttcttct tctgcggcca
     2041 cacgtctggc aaggtttccc tgagagacct ccgtactttt aaggtggaac atgagtttga
     2101 tgccttctca ggaagtctgt cagactttga tgtgcatggc aacctgctag ctgcctgtgg
     2161 cttctccagc cgcctcactg gcctggcctg cgaccgtttc ctcaaggtgt atgatttgcg
     2221 catgatgcgt gccatcacac cacttcaagt acatgtggat cctgccttct tgcgcttcat
     2281 tcctacatat acttctcgtc ttgctatcat ctctcagtca gggcagtgcc aattctgtga
     2341 acccacaggc ctggccaacc cagccgatat ctttcatgtg aatcctgtgg ggcctctgct
     2401 aatgacattt gatgtgtcag ccagcaagca ggctctggcc tttggggatt ctgagggctg
     2461 tgtgcacctc tggactgatt ccccggagcc ttccttcaac ccctactccc gtgagactga
     2521 gtttgctttg ccgtgtctcg tggactcact gcctcctctg gactggagcc aggacctgct
     2581 gcctctttcc ctcatccctg tcccactcac cactgacaca cttctctctg attggcctgc
     2641 tgccaactct gctccagctc ccaggcgagc accacccgtg gatgcagaga ttctgcgcac
     2701 catgaagaag gtgggcttca ttggctatgc gcccaatccc cgcaccaggc tgcgcaatca
     2761 gataccctac agactcaagg agtcagacag tgaatttgac agcttcagcc aggtcactga
     2821 gtcaccagta ggacgagaag aggaaccaca tctccacatg gtttctaaga aataccgcaa
     2881 ggtgaccatc aaatattcca agctagggct ggaggacttt gacttcaaac actacaataa
     2941 gaccttgttt gctggattag agccccacat tcccaacgcc tactgtaact gcatgatcca
     3001 ggtgctctat ttcctggagc ctgtacgctg tctaattcaa aaccaccttt gccagaagga
     3061 gttctgtctg gcatgtgagc tgggcttcct gtttcacatg ttggacctct ctcgtggtga
     3121 cccttgccag ggcaataatt ttcttcgggc attccgtact attcctgagg cctcagccct
     3181 cggtctaatc ctggctgact cagatgaggc ctcaggcaag ggcaatctgg ccaggctcat
     3241 tcagaggtgg aatcgcttca ttctcactca actgcatcaa gatatgcagg agctggaaat
     3301 accacaggct tatcgaggtg ctggaggcag cagcttttgc tcatcggggg actctgttat
     3361 tgggcagctc ttcagctgtg agatggagaa ctgcagcctc tgccgctgtg gcagtgagac
     3421 cgtgcgagcc tcatccactc tgcttttcac actctcctac cctgatggta gcaaaagtga
     3481 taaaactggg aagaactatg actttgctca ggtgctgaag cgaagcatct gcctggacca
     3541 gaatacacag gcctggtgtg acacctgtga aaagtaccag cccacgattc agacccgcaa
     3601 catccgccat ctgccagata ttcttgtcat caattgtgag gtgaacagct caaaagaggc
     3661 tgatttctgg agaatgcagg ctgaggttgc cttcaagatg gcagtaaaga aacacggtgg
     3721 ggaaatctcc aagaacaagg aatttgcttt ggctgattgg aaggaactag ggagtccaga
     3781 gggtgtgctg gtgtgtccct ccattgagga gttgaagaac gtctggcttc ctttctccat
     3841 tcgcatgaag atgaccaaaa acaaagggct ggatgtttgc aattggactg atggggatga
     3901 gatgcagtgg ggcccagcca gggcagagga ggagcatggt gtctatgtgt atgacctgat
     3961 ggctactgtg gtacacatcc tggactcacg cacagggggc agcctggtgg ctcacatcaa
     4021 agttggagag acctaccacc agcgcaagga gggcgttact caccagcagt ggtatctgtt
     4081 caatgacttt cttattgaac ctattgataa gcatgaagct gtgcagtttg acatgaattg
     4141 gaaagtacct gcaatccttt attatgtcaa acggaatctc aattccagat acaacctgaa
     4201 catcaagaac cctattgagg caagtgtctt gctggctgaa gcctcgctgg cacggaagca
     4261 gcggaaaaca catactacct ttattccact gatgctgaat gagatgccac agattgggga
     4321 cctggtgggt ctggatgctg agtttgtcac ccttaatgag gaggaagcag agttacgcag
     4381 tgatggtacc aagtctacca ttaaaccaag ccagatgtca gtagccagga ttacctgtgt
     4441 tcggggccag ggacccaatg agggtatccc cttcattgat gactacatct ctacccagga
     4501 gcaggtggtg gattacttga ctcaatactc gggtataaag cctggtgacc tcgatgccaa
     4561 aatttcctcc aagcacctaa caactctcaa gtctacctac ttaaagcttc gttttctcat
     4621 tgacattgga gtcaagtttg tgggtcatgg cctgcagaag gacttccggg tcgtcaacct
     4681 gatggtgccc aaggaccaag tccttgacac tgtctacctg ttccatatgc cccgaaaacg
     4741 aatgatttcc ctgcgattcc ttgcttggta ctttctggac ctgaagattc aaggggaaac
     4801 ccatgacagt attgaggatg cccgcacagc ccttcagctg taccgaaagt atctggagct
     4861 aagcaaaaat ggcactgagc ctgagtcttt ccacaaggtg ctcaagggtc tttatgagaa
     4921 gggcagaaag atggactgga aggtgcctga gcctgagggc caaacaagtc ccaagaatgc
     4981 agctgtcttc tcctcagtgc tggcgctctg agaattctgc agtcgacggt accgcgggcc
     5041 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     5101 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5161 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5221 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5281 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5341 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5401 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5461 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5521 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5581 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5641 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5701 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5821 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5881 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5941 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6061 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6181 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6241 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6301 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6361 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6421 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     6481 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6541 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6601 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6661 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6721 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6781 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6841 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6901 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6961 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7021 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7081 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     7141 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7201 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     7261 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     7321 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7381 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7441 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7501 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7561 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7621 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7681 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7741 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7801 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8161 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8221 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8281 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8341 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
