LOCUS       sequence-386700-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Ctdnep1_D67E-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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(+)-C-eGFP-Ctdnep1_D67EsiR
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..24)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13..237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      283..711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(370..389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     580..601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(720..740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        725..1054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      905..1040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     967..986
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1121..1915
                     /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(1175..1194)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1785..1804
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2089..2210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2126..2145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2180..2199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2259..2275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2259..2275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2272..2294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2283..2299
                     /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(2307..2313,2314..2331,2332..2337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2352..2373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2490..2507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2661..3249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2741..2760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3420..4211,4212..4280))
                     /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     4043..4062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4281..4385)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4460..4479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4651..5030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5031..5234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5185..5205
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5279..5298
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5279..5297
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5351..5375
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6123..6147)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(6156..6158,6159..6872)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             6156..6869
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(6198..6219)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6459..6478)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6806..6827
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
       61 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
      121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
      181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
      241 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
      301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      961 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     1021 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     1081 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     1141 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1201 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1261 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1321 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1381 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1441 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1501 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1561 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1621 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1681 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1741 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1801 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1861 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1921 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1981 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     2041 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     2101 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2161 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2221 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2281 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2341 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2401 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2461 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2521 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2581 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3421 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3481 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3541 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3601 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3661 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3721 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3781 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3841 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3901 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3961 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4021 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4081 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4141 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4201 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4261 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4381 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4441 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4501 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4561 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4621 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4681 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4741 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4801 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4861 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4921 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4981 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     5041 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5101 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5161 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5221 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     5281 atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtaccga
     5341 gctcggatcc acaagtttgt acaaaaaagc aggctatgat gcggacgcag tgtctgctgg
     5401 ggctgcgcac gttcgtggcc ttcgccgcca agctctggag cttctttatt taccttttgc
     5461 ggaggcagat ccgcacggta attcagtatc agactgttcg atatgatatc ctgcccttat
     5521 ctcctttgtc ccggaatcgc ctagcccagg tgaagaggaa gatcctggtg ctggagctgg
     5581 acgaaaccct gatccattcc catcatgacg gggtgctgag gcctacagtg agacctggga
     5641 cacctcccga cttcatcctc aaggtcgtca tcgataagca tccagtccgg ttttttgtac
     5701 ataagaggcc ccatgtggat ttcttcttag aagtggtaag ccagtggtat gagcttgtgg
     5761 tgttcacagc aagcatggaa atttatggct ctgctgtggc agataaactg gacaacagca
     5821 gaagcattct taagaggaga tactacagac agcactgcac tttggagttg ggcagctaca
     5881 tcaaagacct ctccgtggtc cacagcgacc tgtccagcat cgtgatcctg gacaactccc
     5941 ccggggctta caggagccac ccagacaatg ccatccccat caaatcctgg ttcagtgacc
     6001 ccagtgacac agcccttctc aaccttctcc caatgctgga tgccctcagg ttcactgctg
     6061 atgtccgatc ggtgctgagc cgaaaccttc accaacatag gctctggggc ggcggcggcg
     6121 gcacccagct ttcttgtaca aagtggtgcg gccgcatgag caagggcgag gagctgttca
     6181 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     6241 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     6301 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     6361 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     6421 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     6481 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     6541 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     6601 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6661 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     6721 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     6781 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6841 tcactcacgg catggacgag ctgtacaagt aatctagagg gcccgtttaa acccgct
//
