LOCUS       sequence-183006-        4969 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 4969)
  AUTHORS   Machacek M, Hodgson L, Welch C, Elliott H, Pertz O, Nalbant P, Abell
            A, Johnson GL, Hahn KM, Danuser G
  TITLE     Coordination of Rho GTPase activities during cell protrusion.
  JOURNAL   Nature. 2009 Sep 3. 461(7260):99-103.
  PUBMED    19693013
REFERENCE   2  (bases 1 to 4969)
  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: "Nature. 
            2009 Sep 3. 461(7260):99-103."
COMMENT     Sequence Label: pYpet-PBD
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..308
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     265..285
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(310..414)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      441..896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(528..547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     738..759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    903..1024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(914..933)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     968..987
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1856..1875
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(2213..2416)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2242..2262)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2417..2720
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2895..3483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2975..2994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3812..3859
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3884..3903
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4091..4885)
                     /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             complement(4091..4885)
                     /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(4202..4221)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4812..4831
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4920..4969,1..308))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4934..4969,1..100)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
       61 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
      121 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      181 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      241 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
      301 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      421 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
      541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
      661 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      721 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      781 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      841 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
      901 gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      961 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1021 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1081 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag
     1141 atccggtgga tcccgggccc gcggtaccgt cgactgcaga attcatttct ggctgttgga
     1201 tgtcttcttc gagttgtaaa actccaacac atccagaaca gcctgcgggt ttttcttctg
     1261 ctccgactta gtgatatttg atgtctgaag caagcgggcc cactgctctg gcattcccgt
     1321 aaactcccct gtgacagcat caaaaccgac atgaattgtg tgttcaaaat ctgaagggag
     1381 agaaatctct ggccgctctt tctctttctt tttatttgtt ttatctccag gtaaaatgga
     1441 tccgagctca gatctgagtc cggacttgta cagctcgtcc atgccctcgg tgatgccggc
     1501 ggcggtcagg aactccagca gcaccatgtg gtcccgcttc tcgttggggt ccttgaacag
     1561 ggcgctctgg tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1621 ggtgttctgc tggtagtggt cggccagctg cacgccgccg tcctcgatgt tgtgccggat
     1681 cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatgtaca cgttgtggct
     1741 gttgtagttg tactccagct tgtggcccag gatgttgccg tcctccttga agtcgatgcc
     1801 cttcagctcg atccggttca ccagggtgtc gccctcgaac ttcacctcgg cccgggtctt
     1861 gtagttgccg tcgtccttga agaagatggt ccgctcctgc acgtagccct cgggcatggc
     1921 gctcttgaag aagtcgtgct gcttcatgtg gtcggggtac cgggcgaagc actgcacgcc
     1981 gtagcccagg gtggtcacca gggtgggcca gggcacgggc agcttgccgg tggtgcacag
     2041 cagcttcagg gtcagcttgc cgtaggtggc gtcgccctcg ccctcgccgc tcacgctgaa
     2101 cttgtggccg ttcacgtcgc cgtccagctc caccaggatg ggcaccacgc cggtgaacag
     2161 ctcttcgccc ttgctcacca tggtgctagc ggatctgacg gttcactaaa ccagctctgc
     2221 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     2281 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     2341 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     2401 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     2461 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     2521 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     2581 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     2641 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     2701 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     2761 ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     2821 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2881 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2941 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3001 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3061 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3121 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3181 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3241 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3301 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3361 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3421 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3481 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3541 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3601 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     3661 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     3721 gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     3781 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     3841 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     3901 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     3961 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     4021 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     4081 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4141 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4201 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4261 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     4321 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     4381 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4441 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4501 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4561 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4621 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4681 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     4741 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4801 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4861 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     4921 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaata
//
