LOCUS       sequence-386570-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc-Eps8L2_529-715.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  AUTHORS   Calero-Cuenca FJ, Osorio DS, Carvalho-Marques S, Sridhara SC, 
            Oliveira LM, Jiao Y, Diaz J, Janota CS, Cadot B, Gomes ER
  TITLE     Ctdnep1 and Eps8L2 regulate dorsal actin cables for nuclear 
            positioning during cell migration.
  JOURNAL   Curr Biol. 2021 Apr 12;31(7):1521-1530.e8. doi: 
            10.1016/j.cub.2021.01.007. Epub 2021 Feb 9.
  PUBMED    33567288
REFERENCE   2  (bases 1 to 5318)
  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: 
            "10.1016/j.cub.2021.01.007"; journalName: "Curr Biol"; date: 
            "2021-04-12- 12"; volume: "31"; issue: "7"; pages: "1521-1530.e8"
COMMENT     Sequence Label: pRK5mycGW-Eps8L2_529-715
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    21..155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(58..77)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     112..131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        224..581
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        224..415
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(247..267)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      432..567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(601..618)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(601..617)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(610..632)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      830..1285
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(917..936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1127..1148
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1302..1321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1421..1443
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1481..1499)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1567..1671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1672..1740,1741..2532)
                     /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(1890..1909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3445..3462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3579..3600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3615..3620,3621..3638,3639..3645)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3653..3669
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3658..3680
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3677..3693
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3677..3693
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3722..4101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4102..4305
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4256..4276
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4302..4326
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4536..4554
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4536..4553
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4567..4586
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4635..4664
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4683..4707
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5274..5298)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 aagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca
       61 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      121 ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat
      181 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
      241 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
      301 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
      361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
      421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
      481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
      541 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc
      601 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      661 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      721 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      781 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
      841 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
      901 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
      961 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1021 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     1081 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1141 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     1201 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1261 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     1321 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1381 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1441 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1501 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1561 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1621 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1681 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1741 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1801 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1861 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1921 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1981 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2041 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2101 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2161 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2221 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2581 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3421 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3481 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3541 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     3601 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     3661 ggataacaat ttcacacagg aaacagctat gacatgatta cgaattaatt cgagctcgcc
     3721 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     3781 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     3841 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     3901 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     3961 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4021 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4081 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4141 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4201 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4261 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
     4321 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
     4381 tccagcctcc gcggccggga acggtgcatt ggaacgcgga ttccccgtgc caagagtgac
     4441 gtaagtaccg cctatagagt ctataggccc acccccttgg cttcgttaga acgcggctac
     4501 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaataaca
     4561 tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcac ctcggttcta
     4621 tcgatcccac catggagcag aagctgatca gcgaggagga cctgctcgag ggatccccat
     4681 caacaagttt gtacaaaaaa gcaggctcta tgtggaagct gcgcagccgc agcggccagg
     4741 cggggtacgt gccctgcaac atcctaggcg aggcgcgacc ggaggacgcc ggcgccccgt
     4801 tcgagcaggc cggtcagaag tactggggcc ccgccagccc gacccacaag ctacccccaa
     4861 gcttcccggg caataaggac gaactcatgc agcacatgga cgaggtcaac gacgagctca
     4921 tccgcaagat atcgaatata agggcgcagc cacagaggca cttccgcgtg gagcgcagcc
     4981 agcccgtgag ccagccgctc acctacgagt cgggtccgga cgaggtccgc gcctggctgg
     5041 aagccaaggc cttcagcccg cggatcgtgg agaacctggg catcctgacc gggccgcagc
     5101 tcttctccct caacaaggag gagctgaaga aagtgtgcgg cgaggagggc gtccgcgtgt
     5161 acagccagct caccatgcag aaggccttcc tggagaagca gcaaagtggg tcggagctgg
     5221 aagaactcat gaacaagttt cattccatga atcagaggag gggggaggac agcacccagc
     5281 tttcttgtac aaagtggttg atgggaattc tagattag
//
