LOCUS       40924_44948        4220 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC.Donor.RFP.dATG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4220)
  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 4220)
  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 4220)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4220)
  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..4220
                     /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             566..1237
                     /codon_start=1
                     /label=DsRed-Express
                     /note="rapidly maturing tetrameric variant of DsRed
                     fluorescent protein (Bevis and Glick, 2002)"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHKA
                     LKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHHL
                     FL"
     polyA_signal    complement(1285..1406)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1561..1577)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2051..2155
                     /label=AmpR promoter
     CDS             2156..3013
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3187..3775
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4063..4084
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4099..4129
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4137..4153
                     /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     4161..4177
                     /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 tctaggcctc ctccgaggac gtcatcaagg agttcatgcg
      601 cttcaaggtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
      661 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
      721 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt
      781 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
      841 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
      901 gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg
      961 ccccgtaatg cagaagaaga ctatgggctg ggagccctcc accgagcgcc tgtacccccg
     1021 cgacggcgtg ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta
     1081 cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta
     1141 ctacgtggac tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca
     1201 gtacgagcgc gccgagggcc gccaccacct gttcctgtag cggccgcact taagttatcg
     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 tgtggtaaaa
     1501 tcgaattctg cagtcgacgg taccgcgggc ccgggatccc cgggtaccga gctcgaattc
     1561 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1621 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     1681 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     1741 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     1801 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1861 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1921 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1981 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2041 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2101 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2161 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2221 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2281 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2341 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2401 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2461 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2521 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2581 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2641 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2701 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2761 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2821 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2881 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2941 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3001 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3061 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3181 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3241 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3301 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3661 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3721 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3781 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3841 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3901 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3961 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4021 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4081 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4141 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctggccgc
     4201 acttaagtta cgcgtggatc
//
