LOCUS       sequence-385549-        7614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Myc-Eps8L2-FL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  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 7614)
  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-FL-siR
FEATURES             Location/Qualifiers
     source          1..7614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      915..924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             924..953
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    972..996
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3147..3171)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3203..3220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3209..3433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3479..3907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3566..3585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3776..3797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3916..3936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3921..4250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4101..4236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4163..4182
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4317..5111
                     /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(4371..4390)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4981..5000
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5285..5406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5322..5341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5376..5395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5455..5471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5455..5471)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5468..5490)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5479..5495
                     /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(5503..5509,5510..5527,5528..5533))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5548..5569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5686..5703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5857..6445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5937..5956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6616..7407,7408..7476))
                     /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     7239..7258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7477..7581)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
      901 ttaaacttaa gcttgccacc atggagcaga aactcatctc tgaagaggat ctgggtaccg
      961 agctcggatc cacaagtttg tacaaaaaag caggctctat gagccagtcc ggggccgtga
     1021 gctgctgccc gggtgccacc aatggcagcc tgggccggtc cgacggtgtg gccaagatga
     1081 gccccaagga cctgtttgag cagaggaaga agtattccaa ctccaacgtc atcatgcacg
     1141 agacctcgca gtaccacgtc cagcacctgg ccacattcat catggacaag agcgaagcca
     1201 tcacgtctgt ggacgacgcc atccggaagc tggtgcagct gagctccaag gagaagatct
     1261 ggacccagga gatgctgctg caagtcaatg atcagtcgct gcggctgctg gacatcgagt
     1321 cacaggagga gctggaagac ttcccgctgc ccacggtgca gcgcagccag acggtcctca
     1381 accagctgcg ctacccgtct gtgctgctgc tcgtgtgcca ggactcggag cagagcaagc
     1441 cggatgtcca cttcttccac tgcgatgagg tggaggcaga gctggtgcac gaggacatcg
     1501 agagcgcgtt ggccgactgc cggctgggca agaagatgcg gccgcagacc ctgaagggac
     1561 accaggagaa gattcggcag cggcagtcca tcctgcctcc tccccagggc ccggcgccca
     1621 tccccttcca gcaccgcggc ggggattccc cggaggccaa gaatcgcgtg ggcccgcagg
     1681 tgccactcag cgagccaggt ttccgccgtc gggagtcgca ggaggagccg cgggccgtgc
     1741 tggctcagaa gatagagaag gagacgcaaa ttctcaactg cgccctggac gacatcgagt
     1801 ggtttgtggc ccggctgcag aaggcagccg aggctttcaa gcagctgaac cagcggaaaa
     1861 aggggaagaa gaagggcaag aaggcgccag cagagggcgt cctcacactg cgggcacggc
     1921 ccccctctga gggcgagttc atcgactgct tccagaaaat caagctggcg attaacttgc
     1981 tggcaaagct gcagaagcac atccagaacc ccagcgccgc ggagctcgtg cacttcctct
     2041 tcgggcctct ggacctgatc gtcaacacct gcagtggccc agacatcgca cgctccgtct
     2101 cctgcccact gctctcccga gatgccgtgg acttcctgcg cggccacctg gtccctaagg
     2161 agatgtcgct gtgggagtca ctgggagaga gctggatgcg gccccgttcc gagtggccgc
     2221 gggagccaca ggtgcccctc tacgtgccca agttccacag cggctgggag cctcctgtgg
     2281 atgtgctgca ggaggccccc tgggaggtgg aggggctggc gtctgccccc atcgaggagg
     2341 tgagtccagt gtcgaggcaa agtatccgga actcccagaa gcacagcccc acttcagagc
     2401 ccaccccccc gggggatgcc ctaccaccag tcagctcccc acatactcac aggggctacc
     2461 agccaacacc agccatggcc aaatatgtga aaattctcta tgacttcaca gcccgaaatg
     2521 ccaacgagct atcggtgctc aaggatgagg tcctagaggt gctggaggac ggccggcagt
     2581 ggtggaagct gcgcagccgc agcggccagg cggggtacgt gccctgcaac atcctaggcg
     2641 aggcgcgacc ggaggacgcc ggcgccccgt tcgagcaggc cggtcagaag tactggggcc
     2701 ccgccagccc gacccacaag ctacccccaa gcttcccggg caataaggac gaactcatgc
     2761 agcacatgga cgaggtcaac gacgagctca tccgcaagat atcgaatata agggcgcagc
     2821 cacagaggca cttccgcgtg gagcgcagcc agcccgtgag ccagccgctc acctacgagt
     2881 cgggtccgga cgaggtccgc gcctggctgg aagccaaggc cttcagcccg cggatcgtgg
     2941 agaacctggg catcctgacc gggccgcagc tcttctccct caacaaggag gagctgaaga
     3001 aagtgtgcgg cgaggagggc gtccgcgtgt acagccagct caccatgcag aaggccttcc
     3061 tggagaagca gcaaagtggg tcggagctgg aagaactcat gaacaagttt cattccatga
     3121 atcagaggag gggggaggac agctaaaccc agctttcttg tacaaagtgg ttctagaggg
     3181 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3241 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3301 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3361 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3421 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3481 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3541 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3601 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3661 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3721 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3781 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3841 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3901 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4081 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4141 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4201 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4261 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4321 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4381 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4441 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4501 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4561 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4621 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4681 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4741 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4801 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4861 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4921 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4981 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5041 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5101 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5161 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5221 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5281 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5401 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5461 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5521 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5581 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5641 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5701 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6781 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6901 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7141 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7201 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7441 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7561 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcga
//
