LOCUS       sequence-91659-e        5890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5890)
  AUTHORS   Tanaka T, Serneo FF, Higgins C, Gambello MJ, Wynshaw-Boris A, 
            Gleeson JG
  TITLE     Lis1 and doublecortin function with dynein to mediate coupling of 
            the nucleus to the centrosome in neuronal migration.
  JOURNAL   J Cell Biol. 2004 Jun 7. 165(5):709-21.
  PUBMED    15173193
REFERENCE   2  (bases 1 to 5890)
  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"; journalName: "J Cell 
            Biol. 2004 Jun 7. 165(5):709-21."
COMMENT     Sequence Label: EGFP-DCX
FEATURES             Location/Qualifiers
     source          1..5890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1357..1374
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1384..1401
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2506..2538
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     polyA_signal    2681..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2718..2737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2772..2791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2809..3264)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3158..3177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3291..3395
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3397..3752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3397..3586
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3420..3440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3603..3738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3841..3860)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4450..4469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4768..4787)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4812..4859
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5188..5776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5677..5696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atccatgggc agcagccatc atcatcatca tcacagcagc
     1381 ggcctggtgc cgcgcggcag ccatatggaa cttgattttg gacactttga cgaaagagat
     1441 aagacatcca ggaacatgcg aggctcccgg atgaatgggt tgcctagccc cactcacagc
     1501 gcccactgta gcttctaccg aaccagaacc ttgcaggcac tgagtaatga gaagaaagcc
     1561 aagaaggtac gtttctaccg caatggggac cgctacttca aggggattgt gtacgctgtg
     1621 tcctctgacc gttttcgcag ctttgacgcc ttgctggctg acctgacgcg atctctgtcc
     1681 gacaacatca acctgcctca gggagtgcga tacatttaca ccattgatgg atccaggaag
     1741 atcggaagca tggatgaact ggaggaaggg gaaagctatg tctgttcctc agacaacttc
     1801 tttaaaaagg tggagtacac caagaatgtc aatcccaact ggtctgtcaa cgtaaaaaca
     1861 tctgccaata tgaaagcccc ccagtctttg gctagcagca acagtgcaca ggccagggag
     1921 aacaaggact ttgtgcgccc caagctggtt accatcatcc gcagtggggt gaagcctcgg
     1981 aaggctgtgc gtgtgcttct gaacaagaag acagcccact cttttgagca agtcctcact
     2041 gatatcacag aagccatcaa actggagacc ggggttgtca aaaaactcta cactctggat
     2101 ggaaaacagg taacttgtct ccatgatttc tttggtgatg atgatgtgtt tattgcctgt
     2161 ggtcctgaaa aatttcgcta tgctcaggat gatttttctc tggatgaaaa tgaatgccga
     2221 gtcatgaagg gaaacccatc agccacagct ggcccaaagg catccccaac acctcagaag
     2281 acttcagcca agagccctgg tcctatgcgc cgaagcaagt ctccagctga ctcaggtaac
     2341 gaccaagacg caaacggaac ctccagcagc cagctctcta cccccaagtc taagcagtct
     2401 cccatctcta cgcccaccag tcctggcagc ctccggaagc acaaggacct gtacctgcct
     2461 ctgtccttgg atgactcgga ctcgcttggt gattccatgt aacatatggc tagcatgact
     2521 ggtggacagc aaatgggtcg cggatccgaa ttgatccacc ggatctagat aactgatcat
     2581 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2641 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2761 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2821 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2881 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2941 ttgttccagt ttggaacaag actccactat taaagaacgt ggactccaac gtcaaagggc
     3001 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3061 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3121 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3181 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3241 ttaattcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3361 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3421 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcagggc gatgtatgca
     3481 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct cccagcaggc
     3541 agagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaagtacgc
     3601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3661 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3721 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3781 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3841 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3901 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3961 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4021 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4081 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4141 gcaatgcggc ggctgcatac gcttgatccg gctacctccc attcgaccac caagcgaaac
     4201 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4261 acgaagagca tcagggcgat gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4321 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4381 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4441 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4501 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4561 ttcttgacga gttcttctga gccggactca ggggttcgaa atgaccgacc aagcgacgcc
     4621 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4681 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4741 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4801 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4861 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4921 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4981 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5821 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5881 cgccatgcat
//
