LOCUS       Exported                7643 bp DNA     circular SYN 19-NOV-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7643)
  AUTHORS   Bennardo N, Cheng A, Huang N, Stark JM
  TITLE     Alternative-NHEJ is a mechanistically distinct pathway of mammalian 
            chromosome break repair.
  JOURNAL   PLoS Genet. 2008 Jun 27;4(6):e1000110. doi: 
            10.1371/journal.pgen.1000110.
  PUBMED    18584027
REFERENCE   2  (bases 1 to 7643)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7643)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7643)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
            Genet."; date: "2008-06-27"; pages: "
            10.1371/journal.pgen.1000110"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..658
                     /label=chicken beta-actin promoter
     intron          659..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1683..1702
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1764..1784
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1806..1826
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             2081..2797
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3005..3024)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3070..3125
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3124..3143)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     promoter        complement(3483..3501)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3582..3601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     3701..3723
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3761..3779)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        3847..3951
                     /label=AmpR promoter
     CDS             3952..4809
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4983..5571
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5725..5742
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5829..5847
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5885..6382
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             6401..6997
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        7641..7643
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken
                     beta-actin promoter"
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 gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattaa ttcagatccg
     1741 ccgccactat gggatcaaga tcgccaaaaa agaagagaaa ggtgccgaag aagcatgcag
     1801 caccaccaaa aaaaaaacga aaagtagaag acccacgagg caacaccagc ggcgtgctga
     1861 gcacccccaa ggccaagagg gccaagcacc cccccggcac cgagaagccc aggagcagga
     1921 gccagagcga gcagcccgcc acctgcccca tctgctacgc cgtgatcagg cagagcagga
     1981 acctgaggag gcacctggag ctgaggcact tcgccaagcc cgcctaggga taacagggta
     2041 attagatgac aagcccggcg tggatccacc ggtcgccacc atggtgagca agggcgagga
     2101 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2161 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2221 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2281 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2341 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2401 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2461 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2521 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2581 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2641 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2701 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2761 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     2821 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2881 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaattcact cctcaggtgc
     2941 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     3001 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     3061 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     3121 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     3181 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     3241 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     3301 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     3361 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     3421 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag ctggcgtcga
     3481 gttctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     3541 ccgcgttcta acgacaatat gtccatatgg tgcactctca gtacaatctg ctctgatgcc
     3601 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3661 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3721 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3781 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3841 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3901 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3961 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4021 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4081 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4141 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4201 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4261 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4321 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4381 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4441 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4501 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4561 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4621 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4681 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4741 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4801 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4861 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4921 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4981 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5041 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5101 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5161 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5221 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5281 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5341 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5401 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5461 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5521 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5581 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5641 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5701 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5761 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggtt aacctggctt
     5821 atcgaaatta atacgactca ctatagggag accggcagat ctgatatcat cgatgaattc
     5881 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     5941 ctgggcactt ggcgctacac aagtggcctc tggcctcaca cattccacat ccaccggtag
     6001 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     6061 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     6121 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     6181 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     6241 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     6301 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     6361 ttcctcatct ccgggccttt cgaccgatcc agccgccacc atgaccgagt acaagcccac
     6421 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     6481 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     6541 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     6601 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     6661 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     6721 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6781 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     6841 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     6901 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6961 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     7021 gcgcccgacc gaaaggagcg cacgacccca tggctccgac cgaagccacc cggggcggcc
     7081 ccgccgaccc cgcacccgcc cccgaggccc accgcggggg acacaccgaa cacgccgacc
     7141 ctgctgaaca cgcggcgcag ttcggtgccc aggagcggat cgaaattgat gatctattaa
     7201 acaataaaga tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga
     7261 gaacagagta cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg
     7321 gattagataa atgcctgctc tttactgaag gctctttact attgctttat gataatgttt
     7381 catagttgga tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc
     7441 cactcatgat ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc
     7501 actttgtggt tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga
     7561 gatcagcagc ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat
     7621 tccatcagaa gcttcagctg ctc
//
