LOCUS       40924_44943        4180 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC.Donor.GFP.dATG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4180)
  AUTHORS   Holkers M, de Vries AA, Goncalves MA.
  TITLE     Nonspaced inverted DNA repeats are preferential targets for 
            homology-directed gene repair in mammalian cells
  JOURNAL   Nucleic Acids Res. 40 (5), 1984-1999 (2012)
  PUBMED    22080552
REFERENCE   2  (bases 1 to 4180)
  AUTHORS   Holkers M, de Vries AF, Goncalves MAF.V.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAR-2011) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden, Zuid-Holland 2333 ZC, The 
            Netherlands
REFERENCE   3  (bases 1 to 4180)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4180)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2012"; volume: "40"; issue: "5"; pages: 
            "1984-1999"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAR-2011) Molecular Cell Biology, Leiden University Medical 
            Center, Einthovenweg 20, Leiden, Zuid-Holland 2333 ZC, The 
            Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    148..203
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             567..1250
                     /codon_start=1
                     /label=hrGFP
                     /note="humanized Renilla green fluorescent protein"
                     /translation="VEEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGNQLVQIRVTKGA
                     PLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYEDGGLVEIRSDI
                     NLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVGQVILVYRLNSG
                     KFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAIAQLTSLGKPLG
                     SLHEWV"
     polyA_signal    complement(1285..1406)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1521..1537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2011..2115
                     /label=AmpR promoter
     CDS             2116..2973
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3147..3735
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4023..4044
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4059..4089
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4097..4113
                     /label=lac operator
                     /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)."
     primer_bind     4121..4137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
       61 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
      121 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
      181 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
      241 atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa
      301 caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
      361 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
      421 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
      481 tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag
      541 cccaagctga tcccgggatt tctagagtcg aggagatcat gagcttcaag gtgaacctgg
      601 agggcgtggt gaacaaccac gtgttcacca tggagggctg cggcaagggc aacatcctgt
      661 tcggcaacca gctggtgcag atccgcgtga ccaagggcgc ccccctgccc ttcgccttcg
      721 acatcctgag ccccgccttc cagtacggca accgcacctt caccaagtac cccgaggaca
      781 tcagcgactt cttcatccag agcttccccg ccggcttcgt gtacgagcgc accctgcgct
      841 acgaggacgg cggcctggtg gagatccgca gcgacatcaa cctgatcgag gagatgttcg
      901 tgtaccgcgt ggagtacaag ggccgcaact tccccaacga cggccccgtg atgaagaaga
      961 ccatcaccgg cctgcagccc agcttcgagg tggtgtacat gaacgacggc gtgctggtgg
     1021 gccaggtgat cctggtgtac cgcctgaaca gcggcaagtt ctacagctgc cacatgcgca
     1081 ccctgatgaa gagcaagggc gtggtgaagg acttccccga gtaccacttc atccagcacc
     1141 gcctggagaa gacctacgtg gaggacggcg gcttcgtgga gcagcacgag accgccatcg
     1201 cccagctgac cagcctgggc aagcccctgg gcagcctgca cgagtgggtg taatagctcg
     1261 agcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     1321 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1381 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1441 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggatccc
     1501 cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     1681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     1741 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     1801 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1861 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1921 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1981 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2041 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2101 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2161 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2221 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2281 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2341 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2401 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2461 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2521 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2581 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2641 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2701 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2761 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2821 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2881 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2941 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3361 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3421 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3481 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3541 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3601 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3661 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3721 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3781 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3841 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3901 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3961 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4021 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     4081 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4141 gattacgcca agctggccgc acttaagtta cgcgtggatc
//
