LOCUS       sequence-47079-e        6823 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 6823)
  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 6823)
  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-HsPAN3
FEATURES             Location/Qualifiers
     source          1..6823
                     /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    3611..3732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3648..3667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3702..3721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3739..4194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4088..4107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4221..4325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4327..4684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4327..4518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4350..4370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4535..4670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4719..5513
                     /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             4719..5513
                     /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(4773..4792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5383..5402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5701..5720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5745..5792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6121..6709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6610..6629
                     /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 ctatggatgg aggtgcttta actgatacaa gtctcacaga
     1441 ttcctatttt agcaccagct ttattggagt caatggattt ggaagccctg tagaaacaaa
     1501 atatcccctg atgcagagaa tgactaatag tagcagctcc ccaagccttc taaatgacag
     1561 tgccaagcca tattcagccc atgatcctct aacatcacct gcttcatcct tgtttaatga
     1621 ctttggtgcc ctcaacatct ctcagagacg aaagacacca aatcctactg caagcgagtt
     1681 tattcctaaa ggaggatcaa cctccaggct gagtaacgtg tcccagtcaa atatgtctgc
     1741 cttctctcaa gttttctctc acccatccat gggaagccct gctactgctg gattagcgcc
     1801 aggaatgtcg ttgtctgctg ggtcttcccc tcttcattcc cccaagatta ctccacatac
     1861 ttctcctgct cccagaagaa gaagtcacac tccaaatcca gcaagttaca tggtgccttc
     1921 tagtgcctct acatctgtta ataatcctgt ttctcagact ccgtcttctg gtcaggtgat
     1981 ccaaaaggaa actgttggtg ggacgactta cttctataca gacacaactc cagcaccttt
     2041 gactggaatg gtgtttccaa actatcatat ttatcctcca actgcacctc acgttgctta
     2101 tatgcaaccg aaagcaaacg caccttcctt cttcatggct gatgaactcc gacaggagct
     2161 gatcaacaga catttaataa caatggctca aattgatcaa gcagatatgc cagcagttcc
     2221 tacagaggtt gacagctacc atagcctatt ccctctagaa ccactgccac ctcccaaccg
     2281 gatacagaaa tcaagtaatt ttggatatat tacatcttgc tacaaagctg taaacagcaa
     2341 agatgatctg ccatattgcc ttcggaggat acatggtttt cgtcttgtta acacaaagtg
     2401 catggtgttg gtcgacatgt ggaagaaaat tcaacactca aatatcgtaa ctttgcgtga
     2461 agtatttacc actaaagcat ttgctgagcc ctctcttgtg tttgcacatg atttccatgc
     2521 tggaggagaa actatgatga gcagacactt taatgaccct aatgctgatg cctacttcac
     2581 caagagaaag tggggtcagc acgagggacc attgcccagg cagcatgctg gattattgcc
     2641 agaatctctt atttgggcat atattgtcca actaagttct gcattgcgta ccattcatac
     2701 agcaggtttg gcatgtcgag ttatggatcc aacaaagatt ctgataactg gcaaaacaag
     2761 gttgcgagta aattgtgttg gagtttttga tgttttaaca tttgataaca gtcaaaataa
     2821 taatccattg gcattaatgg cccagtacca gcaagcagat ctgatatcat taggaaaagt
     2881 tgtgttggct ttggcttgca actctttggc aggaattcag cgagagaatt tacagaaagc
     2941 catggaactg gtgacaatca actattcctc tgacctgaag aatctgattt tgtatttgtt
     3001 gactgaccaa aacaggatgc gaagtgtaaa tgacatcatg cccatgattg gtgctcgatt
     3061 ttatactcaa ttggatgctg ctcaaatgag aaatgatgtc atagaggaag accttgcaaa
     3121 ggaggttcaa aatggaagac tgtttaggct cctagcaaaa ttgggaacaa tcaatgagag
     3181 gccggagttt cagaaggatc ccacttggtc agagactgga gaccgttatc tgttgaaact
     3241 ctttagggat catctttttc atcaggtgac agaagcaggt gctccctgga ttgacctcag
     3301 tcatataatt tcttgtctta acaagctaga tgctggtgtg ccagaaaaaa taagcctgat
     3361 ttccagagat gagaagagtg tacttgtggt gacctacagt gacttaaagc gctgctttga
     3421 aaatactttt caagaactga ttgcagctgc aaatggtcag ttgtagggta ccgcgggccc
     3481 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3541 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3601 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3721 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3781 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3841 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3901 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3961 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4021 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4081 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4141 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4261 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4321 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4381 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4621 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4681 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5221 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5701 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5761 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5821 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5881 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5941 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6241 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6781 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
