LOCUS       sequence-386703-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Myc-Eps8L2_1-496.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  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 6957)
  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: pcDNA3.1-Nmyc-Eps8L2_1-496-siR
FEATURES             Location/Qualifiers
     source          1..6957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             926..955
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    974..998
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2492..2516)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2548..2565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2554..2778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2824..3252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2911..2930)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3121..3142
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3261..3281)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3266..3595
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3446..3581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3508..3527
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3662..4456
                     /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(3716..3735)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4326..4345
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4630..4751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4667..4686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4721..4740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4800..4816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4800..4816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4813..4835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4824..4840
                     /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)."
     promoter        complement(join(4848..4854,4855..4872,4873..4878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4893..4914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5031..5048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5202..5790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5282..5301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5961..6752,6753..6821))
                     /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     6584..6603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6822..6926)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggagca gaaactcatc tctgaagagg atctgggtac
      961 cgagctcgga tccacaagtt tgtacaaaaa agcaggctct atgagccagt ccggggccgt
     1021 gagctgctgc ccgggtgcca ccaatggcag cctgggccgg tccgacggtg tggccaagat
     1081 gagccccaag gacctgtttg agcagaggaa gaagtattcc aactccaacg tcatcatgca
     1141 cgagacctcg cagtaccacg tccagcacct ggccacattc atcatggaca agagcgaagc
     1201 catcacgtct gtggacgacg ccatccggaa gctggtgcag ctgagctcca aggagaagat
     1261 ctggacccag gagatgctgc tgcaagtcaa tgatcagtcg ctgcggctgc tggacatcga
     1321 gtcacaggag gagctggaag acttcccgct gcccacggtg cagcgcagcc agacggtcct
     1381 caaccagctg cgctacccgt ctgtgctgct gctcgtgtgc caggactcgg agcagagcaa
     1441 gccggatgtc cacttcttcc actgcgatga ggtggaggca gagctggtgc acgaggacat
     1501 cgagagcgcg ttggccgact gccggctggg caagaagatg cggccgcaga ccctgaaggg
     1561 acaccaggag aagattcggc agcggcagtc catcctgcct cctccccagg gcccggcgcc
     1621 catccccttc cagcaccgcg gcggggattc cccggaggcc aagaatcgcg tgggcccgca
     1681 ggtgccactc agcgagccag gtttccgccg tcgggagtcg caggaggagc cgcgggccgt
     1741 gctggctcag aagatagaga aggagacgca aattctcaac tgcgccctgg acgacatcga
     1801 gtggtttgtg gcccggctgc agaaggcagc cgaggctttc aagcagctga accagcggaa
     1861 aaaggggaag aagaagggca agaaggcgcc agcagagggc gtcctcacac tgcgggcacg
     1921 gcccccctct gagggcgagt tcatcgactg cttccagaaa atcaagctgg cgattaactt
     1981 gctggcaaag ctgcagaagc acatccagaa ccccagcgcc gcggagctcg tgcacttcct
     2041 cttcgggcct ctggacctga tcgtcaacac ctgcagtggc ccagacatcg cacgctccgt
     2101 ctcctgccca ctgctctccc gagatgccgt ggacttcctg cgcggccacc tggtccctaa
     2161 ggagatgtcg ctgtgggagt cactgggaga gagctggatg cggccccgtt ccgagtggcc
     2221 gcgggagcca caggtgcccc tctacgtgcc caagttccac agcggctggg agcctcctgt
     2281 ggatgtgctg caggaggccc cctgggaggt ggaggggctg gcgtctgccc ccatcgagga
     2341 ggtgagtcca gtgtcgaggc aaagtatccg gaactcccag aagcacagcc ccacttcaga
     2401 gcccaccccc ccgggggatg ccctaccacc agtcagctcc ccacatactc acaggggcta
     2461 ccagccaaca ccagccatgg ccaaatatta aacccagctt tcttgtacaa agtggttcta
     2521 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2581 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2641 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2701 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2761 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2821 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2881 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2941 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3001 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3061 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3121 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3181 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3241 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3361 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3421 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3481 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3541 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3601 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3661 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3721 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3781 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3841 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3901 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3961 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4021 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4081 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4141 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4201 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4261 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4321 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4381 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4441 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4501 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4561 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4621 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4681 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4741 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4801 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4861 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4921 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4981 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5041 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5101 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5161 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5641 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6061 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6841 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6901 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
