LOCUS       40924_36288        4232 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR6Kamp-LAP(GFP), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4232)
  AUTHORS   Poser I, Sarov M, Hutchins JR, Heriche JK, Toyoda Y, Pozniakovsky A,
            Weigl D, Nitzsche A, Hegemann B, Bird AW, Pelletier L, Kittler R, 
            Hua S, Naumann R, Augsburg M, Sykora MM, Hofemeister H, Zhang Y, 
            Nasmyth K, White KP, Dietzel S, Mechtler K, Durbin R, Stewart AF, 
            Peters JM, Buchholz F, Hyman AA.
  TITLE     BAC TransgeneOmics: a high-throughput method for exploration of 
            protein function in mammals
  JOURNAL   Nat. Methods 5 (5), 409-415 (2008)
  PUBMED    18391959
REFERENCE   2  (bases 1 to 4232)
  AUTHORS   Hyman AA.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2018) Human Genetics, University of Chicago, 900 E
            57th St, Kcbd 10240, Chicago, IL 60605, USA
REFERENCE   3  (bases 1 to 4232)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4232)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2008"; volume: "5"; issue: "5"; pages: "409-415"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2018) Human Genetics, University of Chicago, 900 E 57th St, 
            Kcbd 10240, Chicago, IL 60605, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..27
                     /label=forward recombineering primer sequence
                     /note="forward recombineering primer sequence"
     CDS             28..48
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    61..105
                     /label=S-peptide
                     /note="S-peptide"
     CDS             61..105
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             145..168
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             196..912
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    953..1538
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      1541..1546
                     /regulatory_class="minus_35_signal"
     regulatory      1561..1569
                     /regulatory_class="minus_10_signal"
     CDS             1620..2420
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     oriT            complement(2457..2566)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2725..3113
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        3207..3311
                     /label=AmpR promoter
     CDS             3312..4169
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 gattatgata ttccaactac tgcaagcgag aatctttatt ttcagggcgc cgccaaattc
       61 aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg acagcggagg tggaggttcg
      121 agcggtccct cgggttcgtc gagcctggaa gttctgttcc aggggcccct gtcgtcgagc
      181 ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt tcaccggggt ggtgcccatc
      241 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      301 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      361 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      421 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      481 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      541 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      601 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      661 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      721 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      781 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
      841 cgcgatcaca tggtccttaa ggagttcgtg accgccgccg ggatcactct cggcatggac
      901 gagctgtaca agtaatgatc tagagtcgag ttaattaaga attccgcccc cccccctctc
      961 cctccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     1021 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     1081 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaggg
     1141 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     1201 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     1261 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1321 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1381 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     1441 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     1501 tttcctttga aaaacacgat gataatatgg ccacaacctg ttacattgca caagataaaa
     1561 atatatcatc acgaacagta aaactgtctg cttacataaa cagtaataca aggggtgtta
     1621 tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     1681 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     1741 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     1801 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     1861 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     1921 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     1981 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2041 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2101 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     2161 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     2221 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     2281 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     2341 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     2401 gccttcttga cgagttcttc tgactcgagt tgccccatct gttacgccgg cggtagccgg
     2461 ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa
     2521 gaaggaacac ccactcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg
     2581 gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa
     2641 ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaac
     2701 ggccacgatg cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac
     2761 gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg
     2821 tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac gtactaagct
     2881 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
     2941 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
     3001 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
     3061 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat
     3121 tagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggatagacg
     3181 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3241 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3301 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3361 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3421 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3481 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3541 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3601 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3661 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3721 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacgtg ggggatcacg
     3781 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccgaac gacgagcgtg
     3841 acgccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3901 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3961 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4021 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4081 tagttatcta cacgacgggg agtcaggcaa ctatgggtga acgaaataga cagatcgctg
     4141 agataggtgc ctcactgatt aagcattggt aacccagccc gcctaatgag cgggcttttt
     4201 tttgaacaaa aggctgcagc tggactttta ga
//
