LOCUS       sequence-385550-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Ctdnep1WT-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+CeGFP-Ctdnep1WTsiR
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        480..683
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     634..654
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     728..747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1572..1596)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1605..1607,1608..2321)
                     /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             1605..2318
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1647..1668)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1908..1927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2255..2276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2353..2370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2359..2583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2629..3057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2926..2947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3066..3086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3071..3400
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3251..3386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3313..3332
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3467..4261
                     /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(3521..3540)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4131..4150
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4435..4556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4472..4491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4526..4545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4605..4621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4605..4621)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4618..4640)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4629..4645
                     /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(4653..4659,4660..4677,4678..4683))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4698..4719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4836..4853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5007..5595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5087..5106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5766..6557,6558..6626))
                     /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     6389..6408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6627..6731)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6806..6825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
       61 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      121 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      661 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      721 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta aacttaagct
      781 tggtaccgag ctcggatcca caagtttgta caaaaaagca ggctatgatg cggacgcagt
      841 gtctgctggg gctgcgcacg ttcgtggcct tcgccgccaa gctctggagc ttctttattt
      901 accttttgcg gaggcagatc cgcacggtaa ttcagtatca gactgttcga tatgatatcc
      961 tgcccttatc tcctttgtcc cggaatcgcc tagcccaggt gaagaggaag atcctggtgc
     1021 tggacctgga cgaaaccctg atccattccc atcatgacgg ggtgctgagg cctacagtga
     1081 gacctgggac acctcccgac ttcatcctca aggtcgtcat cgataagcat ccagtccggt
     1141 tttttgtaca taagaggccc catgtggatt tcttcttaga agtggtaagc cagtggtatg
     1201 agcttgtggt gttcacagca agcatggaaa tttatggctc tgctgtggca gataaactgg
     1261 acaacagcag aagcattctt aagaggagat actacagaca gcactgcact ttggagttgg
     1321 gcagctacat caaagacctc tccgtggtcc acagcgacct gtccagcatc gtgatcctgg
     1381 acaactcccc cggggcttac aggagccacc cagacaatgc catccccatc aaatcctggt
     1441 tcagtgaccc cagtgacaca gcccttctca accttctccc aatgctggat gccctcaggt
     1501 tcactgctga tgtccgatcg gtgctgagcc gaaaccttca ccaacatagg ctctggggcg
     1561 gcggcggcgg cacccagctt tcttgtacaa agtggtgcgg ccgcatgagc aagggcgagg
     1621 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1681 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1741 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1801 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1861 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1921 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1981 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2041 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2101 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2161 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2221 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2281 ccgccgggat cactcacggc atggacgagc tgtacaagta atctagaggg cccgtttaaa
     2341 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2521 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2581 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3061 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3121 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3241 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3301 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3361 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3421 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3481 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3541 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3601 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3661 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3721 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3781 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3841 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3901 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3961 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4021 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4081 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4141 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4201 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4261 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4321 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4381 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4561 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4621 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4681 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4741 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4801 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4861 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4921 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4981 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5041 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5101 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5161 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5221 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5281 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5341 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5401 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5461 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5521 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5581 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5641 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5701 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5761 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5821 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5881 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5941 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6001 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6061 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6121 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6181 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6241 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6301 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6361 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6421 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6481 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6541 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6601 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6661 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6721 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     6781 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6841 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaag
//
