LOCUS       sequence-9395-ex        8436 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 8436)
  AUTHORS   Landthaler M, Gaidatzis D, Rothballer A, Chen PY, Soll SJ, Dinic L, 
            Ojo T, Hafner M, Zavolan M, Tuschl T
  TITLE     Molecular characterization of human Argonaute-containing 
            ribonucleoprotein complexes and their bound target mRNAs.
  JOURNAL   RNA. 2008 Dec;14(12):2580-96. doi: 10.1261/rna.1351608. Epub 2008 
            Oct 31.
  PUBMED    18978028
REFERENCE   2  (bases 1 to 8436)
  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"; doi: 
            "10.1261/rna.1351608"; journalName: "RNA"; date: "2008-12"; volume: 
            "14"; issue: "12"; pages: "2580-96"
COMMENT     Sequence Label: pDESTmycEIF2C1
FEATURES             Location/Qualifiers
     source          1..8436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             639..668
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    678..702
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3362..3381)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3363..3381)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3392..3409)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3392..3408)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3401..3423)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3813..3947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3850..3869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3904..3923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4073..4528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4160..4179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4370..4391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4647..5004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4647..4838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4670..4690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4855..4990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5055..5849
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5913..5961
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6002..6021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6121..6143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6181..6199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6267..6371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6372..6440,6441..7232)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6590..6609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7403..7991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7892..7911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8145..8162
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggta ccaccatgga acaaaaactt atttctgaag
      661 aagatctgtc tagatcaaca agtttgtaca aaaaagcagg ctccaccatg gaagcgggac
      721 cctcgggagc agctgcgggc gcttacctgc cccccctgca gcaggtgttc caggcacctc
      781 gccggcctgg cattggcact gtggggaaac caatcaagct cctggccaat tactttgagg
      841 tggacatccc taagatcgac gtgtaccact acgaggtgga catcaagccg gataagtgtc
      901 cccgtagagt caaccgggaa gtggtggaat acatggtcca gcatttcaag cctcagatct
      961 ttggtgatcg caagcctgtg tatgatggaa agaagaacat ttacactgtc acagcactgc
     1021 ccattggcaa cgaacgggtc gactttgagg tgacaatccc tggggaaggg aaggatcgaa
     1081 tctttaaggt ctccatcaag tggctagcca ttgtgagctg gcgaatgctg catgaggccc
     1141 tggtcagcgg ccagatccct gttcccttgg agtctgtgca agccctggat gtggccatga
     1201 ggcacctggc atccatgagg tacacccctg tgggccgctc cttcttctca ccgcctgagg
     1261 gctactacca cccgctgggg ggtgggcgcg aggtctggtt cggctttcac cagtctgtgc
     1321 gccctgccat gtggaagatg atgctcaaca ttgatgtctc agccactgcc ttttataagg
     1381 cacagccagt gattgagttc atgtgtgagg tgctggacat caggaacata gatgagcagc
     1441 ccaagcccct cacggactct cagcgcgttc gcttcaccaa ggagatcaag ggcctgaagg
     1501 tggaagtcac ccactgtgga cagatgaaga ggaagtaccg cgtgtgtaat gttacccgtc
     1561 gccctgctag ccatcagaca ttccccttac agctggagag tggacagact gtggagtgca
     1621 cagtggcaca gtatttcaag cagaaatata accttcagct caagtatccc catctgccct
     1681 gcctacaagt tggccaggaa caaaagcata cctaccttcc cctagaggtc tgtaacattg
     1741 tggctgggca gcgctgtatt aaaaagctga ccgacaacca gacctcgacc atgataaagg
     1801 ccacagctag atccgctcca gacagacagg aggagatcag tcgcctgatg aagaatgcca
     1861 gctacaactt agatccctac atccaggaat ttgggatcaa agtgaaggat gacatgacgg
     1921 aggtgacagg gcgagtgctg ccggcgccca tcttgcagta cggcggccgg aaccgggcca
     1981 ttgccacacc caatcagggt gtctgggaca tgcgggggaa acagttctac aatgggattg
     2041 agatcaaagt ctgggccatc gcctgcttcg caccccaaaa acagtgtcga gaagaggtgc
     2101 tcaagaactt cacagaccag ctgcggaaga tttccaagga tgcggggatg cctatccagg
     2161 gtcaaccttg tttctgcaaa tatgcacagg gggcagacag cgtggagcct atgttccggc
     2221 atctcaagaa cacctactca gggctgcagc tcattattgt catcctgcca gggaagacgc
     2281 cggtgtatgc tgaggtgaaa cgtgtcggag atacactctt gggaatggct acgcagtgtg
     2341 tgcaggtgaa gaacgtggtc aagacctcac ctcagactct gtccaacctc tgcctcaaga
     2401 tcaatgtcaa acttggtggc attaacaaca tcctagtccc acaccagcgc tctgccgttt
     2461 ttcaacagcc agtgatattc ctgggagcag atgttacaca ccccccagca ggggatggga
     2521 aaaaaccttc tatcacagca gtggtaggca gtatggatgc ccaccccagc cgatactgtg
     2581 ctactgtgcg ggtacagcga ccacggcaag agatcattga agacttgtcc tacatggtgc
     2641 gtgagctcct catccaattc tacaagtcca cccgtttcaa gcctacccgc atcatcttct
     2701 accgagatgg ggtgcctgaa ggccagctac cccagatact ccactatgag ctactggcca
     2761 ttcgtgatgc ctgcatcaaa ctggaaaagg actaccagcc tgggatcact tatattgtgg
     2821 tgcagaaacg ccatcacacc cgccttttct gtgctgacaa gaatgagcga attgggaaga
     2881 gtggtaacat cccagctggg accacagtgg acaccaacat cacccaccca tttgagtttg
     2941 acttctatct gtgcagccac gcaggcatcc agggcaccag ccgaccatcc cattactatg
     3001 ttctttggga tgacaaccgt ttcacagcag atgagctcca gatcctgacg taccagctgt
     3061 gccacactta cgtacgatgc acacgctctg tctctatccc agcacctgcc tactatgccc
     3121 gcctggtggc tttccgggca cgataccacc tggtggacaa ggagcatgac agtggagagg
     3181 ggagccacat atcggggcag agcaatgggc gggaccccca ggccctggcc aaagccgtgc
     3241 aggttcacca ggatactctg cgcaccatgt acttcgcttg acggaattcg cggccgcact
     3301 cgagatatct agacccagct ttaaagtggt tgatcgcgtg catgcgacgt catagctctc
     3361 tccctatagt gagtcgtatt ataagctagg cactggccgt cgttttacaa cgtcgtgact
     3421 gggaaaactg ctagcttggg atctttgtga aggaacctta cttctgtggt gtgacataat
     3481 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     3541 aatgtgttaa actagctgca tatgcttgct gcttgagagt tttgcttact gagtatgatt
     3601 tatgaaaata ttatacacag gagctagtga ttctaattgt ttgtgtattt tagattcaca
     3661 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     3721 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3781 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3841 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3901 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat cctgcattaa
     3961 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctggcgtaa tagcgaagag
     4021 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     4081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4201 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4261 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4381 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4441 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaatatt taacgcgaat
     4501 tttaacaaaa tattaacgtt tacaatttcg cctgatgcgg tattttctcc ttacgcatct
     4561 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     4621 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     4681 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4741 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4801 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4861 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4921 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4981 tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta
     5041 aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5881 agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc
     5941 tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca
     6001 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6061 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6121 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6181 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6241 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6301 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6361 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6421 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6481 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6541 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6601 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6661 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6721 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6781 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6841 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6901 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6961 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7021 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7081 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7141 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7201 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7261 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7321 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7381 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7441 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7501 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     7561 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7621 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7681 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7741 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     7801 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7861 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7921 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7981 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8041 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     8101 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     8161 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     8221 aatgcagagc ttgcaattcg cgcgtttttc aatattattg aagcatttat cagggttatt
     8281 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8341 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     8401 cctataaaaa taggcgtagt acgaggccct ttcact
//
