LOCUS       sequence-11456-e        8665 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 8665)
  AUTHORS   Singla V, Romaguera-Ros M, Garcia-Verdugo JM, Reiter JF
  TITLE     Ofd1, a human disease gene, regulates the length and distal 
            structure of centrioles.
  JOURNAL   Dev Cell. 2010 Mar 16. 18(3):410-24.
  PUBMED    20230748
REFERENCE   2  (bases 1 to 8665)
  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: "Dev Cell. 
            2010 Mar 16. 18(3):410-24."
COMMENT     Sequence Label: pIRES2-eGFP-Ofd1-Myc-G139S
FEATURES             Location/Qualifiers
     source          1..8665
                     /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"
     CDS             3943..3972
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    join(4024..4598,4599..4601,4602..4610)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4191..4208)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4418..4437
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4611..4613,4614..4616,4617..5330)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4611..4613,4614..4616,4617..5327)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4656..4677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4917..4936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5264..5285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5453..5574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5490..5509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5544..5563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5581..6036)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5718..5739)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5930..5949
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6063..6167
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6169..6526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6169..6360
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6192..6212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6377..6512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6561..7355
                     /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             6561..7355
                     /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(6615..6634)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7225..7244
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7543..7562)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7587..7634
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7963..8551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8452..8471
                     /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 ccggactcag atctcgagct caagcttcga attatctgca gaattcgccc tttccgtctt
      661 tattccgtcc agagtccctc ctggctcgcg gacccgccca cccttcccat accgtgggag
      721 gaagcgcggc tgcttgacga gttaggggct gaatgtttag ctctcttgct cccctaggtc
      781 cccagagaca agctggattc cgaggtcaag ggctccctct cctcctgcgc cccggcccgg
      841 gccgaggagt ccgaccgtgg aggggcgagc tcgggaggaa agcagatgag gatggctcag
      901 tccaacatgc cccacaagtc tgatgtgttg agtcaagatg aactgcgcaa aaagctatac
      961 cagacgttta aggatcgggg tgtactggac acacttcaga cccaactccg aaaccagcta
     1021 attcatgaat tgatgcatcc tgtattgagt ggagaagtga agccgccatc catttcagtg
     1081 gaagggagtg ccctcctaat aggggcttct aactcgctgg tggcagacca cttacaaaga
     1141 tgtggctatg aatattcact ctctgttttc tttccagaaa gtggtttggc aaaagaaaag
     1201 atatttacta tgcaggatct actacaactc attcggatta acccctcatc cagtctctat
     1261 aaatcattga tttcaggatt tgataaagaa aataaaaaaa gttttcttat gagtttctta
     1321 aaggagttgg cagaatatta tcaagctaaa gagagctgtg atgcggaaac tcagacaagc
     1381 accacatttc ccagccaagt gtccctagcc gagaagtttc agcttattga cgctcaattt
     1441 gcagatggtt ttcctcaccg ttccaagcta gaatctttag agacaaagtt aaacgagtat
     1501 aagaaagaag tacaacacca gcttcaggta gaaatgtgtc ataagctgaa gtattttaga
     1561 gaagctgaaa taacaaaagt taagatggaa gagagaagaa agtatgaaaa ggaattggct
     1621 gagttccaga atgaatttga aagaacttgt caagcaaaaa atgaagccct catttctcag
     1681 gagaagaatt ctctggaaag aattaaaaaa caccgagaga tggaatcaaa agaaatttat
     1741 gctcaaagac agcttctact aaatgatata gctttgctta gaggtagaga agcagagctg
     1801 aaggagcgaa tcgaaacttt tgaattgacc cagaagctcc aagaagaaaa aattaaaagt
     1861 gaggctgagg cacttgagag acgggaacag aatctgaaga atattgaaga cacctatgat
     1921 cagaaactca aaactgaact gctaaagtac caactggagc taaaggatga ctacatcact
     1981 aggacaaata aactcctgga agaggaaagg aagaataaag aaaaaactat tcatttgcaa
     2041 gaggagctca cagttattaa ttcaaaaaag gaagaactca gtaaatctgt taagcatatg
     2101 aaagaagttg agctggaact ggagtctgtg aaagctcagt ttttagcaat atcaaaacaa
     2161 aaccacttgc tgaatgaaaa ggtgagagag atgagtgatt actcacaact gaaagaagag
     2221 aaagtggagc ttcaggcaca aaataaacta cttaaactgc aactggaaga gactaggaat
     2281 gaaaacttac gactcctaga ccgcataact cagccacctc ctgagcttgt gatttttcag
     2341 aaggaattac agaaaacaga aaaggctatg gaactggagc acaaggactt tgaaactcat
     2401 aggcaggctc ttgagaaaca gctgcagagt gaaattgaaa attctgcaca gctaaggacc
     2461 cagatagcag aatatgatgc ttctgtcaaa aggttaactg ttcaggtggc tgagttaaag
     2521 tcacaactaa aacaaactca gatagcccta gagaatgaag tgtaccgaaa tccaaaacac
     2581 tctctgattc attccctgag tggcctccta ttgagcggca agatggcacc ccacagtgaa
     2641 gataaaagtg gggatttctt gaatgtaccc cttgaacaga acaaggttat agcgggtgca
     2701 gttatgtcaa gggtcccacc ttatgtaaat acagcaacag aagctagttc cccagagtct
     2761 gactttgagt ttatagccag tagtactaaa gccaaagtaa gagagctaga gcaagaggct
     2821 gaacgcttgg aaaaggcatt cagaacttac tatcaaagag ctactcagaa tccttctacg
     2881 agccctcagc cggcaaagag tcctccatct gtaaactcgg tggcagctct cagaagcata
     2941 gcttccagtt ccatggacag acctgtttct gcagaggaca gagttgtctc ggaacagcct
     3001 cttggggaca tgctgaagga agaaatgagt gatatgtcca aagcattcat gggcagtgtg
     3061 gtttcacggc cacgcaggac ctcttcctcc acacgcctct cctctacacc ccatccaaaa
     3121 tcgagaagaa gtcttgataa tgaaatgtat ctggaaggtc tcgggagatt acacatgact
     3181 tcctccagcc ctcttcttga cagagtttct gcatcacctg ctgcttcccc tagtccttgt
     3241 cctgagagaa cagcccaggc gtcacctgtt ccatctaggc acagcttctc tggtcttcca
     3301 gagcaaaatg cctgtttata tcagagacaa actgaaacgc aagataaaag tgagctttca
     3361 aatgtggaca agcaatcttt gaaagatgag aaatttgaac cacctttcag atggaacaag
     3421 acagaacagt ttgaagcaga aggactccac cctgctggtg acatgcctgg aatagatttt
     3481 gctgtggcta ctcagtcttc aagactcatt tcatacgact acccaagtgc tgttcagagc
     3541 caaactggag aacaagacga acaggagcta tgggagcttc acatgaaaga acgcaggcag
     3601 agagaagaac aaaggcataa tgaacggcaa gaagctttag agagggaaag aagagagctg
     3661 ggaaaactag agcaggagcg gaggatgatt gaagagtcac tgaagatgga gatggaagaa
     3721 gaattagaaa agagtgttca agaccagaaa gacaaatcag ctcattgtga aaatacttta
     3781 gaaaaataca tgaaaattat ccagcagaga caggaagaat caaatgcaga taagagctcc
     3841 aaaaaatcgg gcaaagaatg ctcactagtg gacatgatga tgcctagtga caaggatgaa
     3901 agttctccag gattttctca tgaagaacca gatgacatgt gggagcaaaa gctcatttct
     3961 gaagaggact tgtaaaaggg cgaattccag cacactggcg gccgttacta gtggatccat
     4021 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     4081 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4141 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     4201 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     4261 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     4321 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     4381 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     4441 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     4501 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     4561 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca
     4621 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4681 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4741 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4801 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     4861 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4921 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4981 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     5041 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     5101 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     5161 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     5221 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     5281 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     5341 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     5401 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     5461 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5521 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     5581 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5641 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5701 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5761 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5821 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5881 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5941 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6001 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     6061 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     6181 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     6541 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     6601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     6661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6721 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     6781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     7021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     7081 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     7141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     7201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     7261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     7321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     7381 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     7441 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     7501 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     7561 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     7621 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     7681 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     7741 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     7801 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     8041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8101 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8641 ggataaccgt attaccgcca tgcat
//
