LOCUS       sequence-200125-        7373 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 7373)
  AUTHORS   Tulu US, Rusan NM, Wadsworth P
  TITLE     Peripheral, non-centrosome-associated microtubules contribute to 
            spindle formation in centrosome-containing cells.
  JOURNAL   Curr Biol. 2003 Oct 28. 13(21):1894-9.
  PUBMED    14588246
REFERENCE   2  (bases 1 to 7373)
  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: "Curr Biol. 
            2003 Oct 28. 13(21):1894-9."
COMMENT     Sequence Label: pIRESneo-PA-GFP-alpha Tubulin
FEATURES             Location/Qualifiers
     source          1..7373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      709..718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(715..717,718..720,721..1431)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(760..781)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1021..1040)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1368..1389
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          2880..3109
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3092..3111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3182..3756,3757..3759,3760..3768)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3349..3366)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3576..3595
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3791..4594
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3854..3873)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4464..4483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4626..4643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4632..4856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4903..4922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4982..4998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4982..4998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4995..5017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5006..5022
                     /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(5030..5036,5037..5054,5055..5060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5075..5096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5213..5230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5384..5972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5464..5483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6143..6934,6935..7003))
                     /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     6766..6785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7004..7108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7183..7202)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgat atctgcggcc tagctagcgc taccggtcgc caccatggtg
      721 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      781 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      841 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      901 accaccttca gctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      961 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1021 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1081 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1141 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1201 aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1261 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1321 agccaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1381 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1441 agatctcgag tgcgtgagtg catctccatc cacgttggcc aggctggtgt ccagattggc
     1501 aatgcctgct gggagctcta ctgcctggaa cacggcatcc agcccgatgg ccagatgcca
     1561 agtgacaaga ccattggggg aggagatgac tccttcaaca ccttcttcag tgagacgggc
     1621 gctggcaagc acgtgccccg ggctgtgttt gtagacttgg aacccacagt cattgatgaa
     1681 gttcgcactg gcacctaccg ccagctcttc caccctgagc agctcatcac aggcaaggaa
     1741 gatgctgcca ataactatgc ccgagggcac tacaccattg gcaaggagat cattgacctt
     1801 gtgttggacc gaattcgcaa gctggctgac cagtgcaccg gtcttcaggg cttcttggtt
     1861 ttccacagct ttggtggggg aactggttct gggttcacct ccctgctcat ggaacgtctc
     1921 tcagttgatt atggcaagaa gtccaagctg gagttctcca tttacccagc accccaggtt
     1981 tccacagctg tagttgagcc ctacaactcc atcctcacca cccacaccac cctggagcac
     2041 tctgattgtg ccttcatggt agacaatgag gccatctatg acatctgtcg tagaaacctc
     2101 gatatcgagc gcccaaccta cactaacctt aaccgcctta ttagccagat tgtgtcctcc
     2161 atcactgctt ccctgagatt tgatggagcc ctgaatgttg acctgacaga attccagacc
     2221 aacctggtgc cctacccccg catccacttc cctctggcca catatgcccc tgtcatctct
     2281 gctgagaaag cctaccatga acagctttct gtagcagaga tcaccaatgc ttgctttgag
     2341 ccagccaacc agatggtgaa atgtgaccct cgccatggta aatacatggc ttgctgcctg
     2401 ttgtaccgtg gtgacgtggt tcccaaagat gtcaatgctg ccattgccac catcaaaacc
     2461 aagcgcagca tccagtttgt ggattggtgc cccactggct tcaaggttgg catcaactac
     2521 cagcctccca ctgtggtgcc tggtggagac ctggccaagg tacagagagc tgtgtgcatg
     2581 ctgagcaaca ccacagccat tgctgaggcc tgggctcgcc tggaccacaa gtttgacctg
     2641 atgtatgcca agcgtgcctt tgttcactgg tacgtgggtg aggggatgga ggaaggcgag
     2701 ttttcagagg cccgtgaaga tatggctgcc cttgagaagg attatgagga ggttggtgtg
     2761 gattctgttg aaggagaggg tgaggaagaa ggagaggaat actaaggatc cgcggccgca
     2821 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     2881 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     2941 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     3001 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     3061 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     3121 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc
     3181 ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     3241 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     3301 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     3361 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     3421 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     3481 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     3541 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     3601 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     3661 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3721 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg ggataattcc
     3781 tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc
     3841 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3901 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3961 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4021 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4081 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4141 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4201 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4261 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4321 caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc
     4381 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4441 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4501 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4561 cgccttctat cgccttcttg acgagttctt ctgaggggat caattctcta gagctcgctg
     4621 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4681 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4741 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4801 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4861 tgaggcggaa agaaccagct ggggctcgag tgcattctag ttgtggtttg tccaaactca
     4921 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4981 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5041 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7141 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7201 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7261 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7321 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
