LOCUS       sequence-133084-        6678 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 6678)
  AUTHORS   Geraldo S, Khanzada UK, Parsons M, Chilton JK, Gordon-Weeks PR
  TITLE     Targeting of the F-actin-binding protein drebrin by the microtubule 
            plus-tip protein EB3 is required for neuritogenesis.
  JOURNAL   Nat Cell Biol. 2008 Oct;10(10):1181-9. Epub 2008 Sep 21.
  PUBMED    18806788
REFERENCE   2  (bases 1 to 6678)
  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: "Nat Cell 
            Biol."; date: "2008-10"; volume: "10(10)"; pages: "1181-9. Epub 2008
            Sep 21"
COMMENT     Sequence Label: drebrin-YFP
FEATURES             Location/Qualifiers
     source          1..6678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2573..2582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2579..2581,2582..2584,2585..3298)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2624..2645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2885..2904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3232..3253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3421..3542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3458..3477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3512..3531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3549..4004)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3686..3707)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3898..3917
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4031..4135
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4137..4494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4137..4328
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4160..4180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4345..4480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4529..5323
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4529..5323
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4583..4602)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5193..5212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5511..5530)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5555..5602
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5931..6519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6420..6439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattca tggccggcgt
      601 cagcttcagc ggccaccgcc tggagctgct ggcggcttac gaggaggtga tccgagagga
      661 gagcgcggcc gactgggctc tgtacacata tgaagatggc tccgatgacc tcaagcttgc
      721 agcatcagga gaagggggct tgcaggagct ttcgggacac tttgagaacc agaaggtgat
      781 gtacggcttc tgcagtgtca aggactccca agctgctctg ccaaaatacg tgctcatcaa
      841 ctgggtgggc gaagatgtgc ctgatgcccg caagtgcgct tgtgccagcc acgtggctaa
      901 ggtggcggag ttcttccagg gtgtcgacgt gatcgtgaac gccagcagcg tggaagacat
      961 agacgcgggt gccatcgggc agcggctctc taacgggctg gcgcgactct ccagccctgt
     1021 gctgcaccga ctgcggctgc gagaggatga gaacgcagag cccgtgggca ccacctacca
     1081 gaagacggat gcagctgtgg aaatgaagcg gattaaccga gagcagttct gggagcaggc
     1141 caagaaggaa gaagagctgc ggaaggagga ggagcggaag aaggccctgg atgagaggct
     1201 caggttcgag caggagcgga tggagcagga gcggcaggag caagaggagc gcgagcggcg
     1261 ctaccgggag cgggagcagc agatcgagga gcacaggagg aaacagcaga ctttagaagc
     1321 ggaagaggcc aagaggcggt tgaaggagca gtctatcttt ggtgaccatc gggatgagga
     1381 ggaagagacc cacatgaaga agtcagagtc ggaggtggag gaggcagcag ctattattgc
     1441 ccagcggcct gacaacccaa gggagttctt caagcagcag gaaagagtcg catcggcctc
     1501 tgcgggcagc tgtgatgtac cctcgccctt caaccatcga ccaggcagcc acctggacag
     1561 ccaccggagg atggcgccca ctcccatccc cacgcggagc ccgtctgact ccagcaccgc
     1621 ctccacccct gtcgctgagc agatagagcg ggccctggat gaggtcacct cctcgcagcc
     1681 tccaccactg ccaccgccac ccccaccagc ccaagagacc caggagccca gccccatcct
     1741 agacagtgag gagaccagag cagcagcccc tcaggcctgg gccggcccca tggaggagcc
     1801 ccctcaggca caggcgcctc cccgggggcc aggcagccct gcagaggact tgatgttcat
     1861 ggagtctgca gagcaggctg tcctggctgc tcccgtggag cctgccacag ctgacgccac
     1921 ggagatccac gatgcagctg acaccattga aactgacact gccactgctg acaccactgt
     1981 tgccaacaac gtaccccccg ccgccaccag cctcattgac ctatggcctg gcaacgggga
     2041 aggggcctcc acactccagg gtgagcccag ggcccccacg ccaccctcgg gtactgaggt
     2101 caccctggca gaggtgcccc tgctggatga ggtggctccg gagccactgc tgccagcagg
     2161 cgaaggctgt gccacccttc tcaactttga tgagctgcct gagccgccag ccaccttctg
     2221 tgacccagag gaagtggaag gggagcccct ggctgccccc cagaccccaa ctctgccctc
     2281 agcccttgag gagctggagc aagagcagga gccggagccc cacctgctaa ccaatggcga
     2341 gaccacccag aaggagggga cccaggccag tgaggggtac ttcagtcaat cacaggagga
     2401 ggagtttgcc caatcggaag agctctgtgc caaggctccg cctcctgtgt tctacaacaa
     2461 gcctccagag atcgacatca catgctggga tgcagaccca gttccagaag aggaggaggg
     2521 cttcgagggt ggtgatttct cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
     2581 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2641 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2701 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2761 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     2821 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2881 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2941 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3001 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3061 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3121 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3181 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3241 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     3301 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3361 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3541 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3601 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3661 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3721 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3781 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3841 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3901 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3961 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     4141 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4501 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4561 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4621 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4681 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4741 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4801 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4861 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4921 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4981 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5041 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5101 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5161 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5221 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5281 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5341 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5401 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5461 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5521 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5581 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5641 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5701 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5761 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5821 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5881 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5941 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6001 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6061 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6121 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6181 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6241 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6301 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6361 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6421 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6481 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6541 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6601 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6661 gtcattagtt catagccc
//
