LOCUS       40924_43548        4491 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Donor vector pTKIP-hygro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Site-specific chromosomal integration of large synthetic constructs
  JOURNAL   Nucleic Acids Res. 38 (6), E92 (2010)
  PUBMED    20047970
REFERENCE   2  (bases 1 to 4491)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2009) Molecular Biology, Princeton University, 
            Washington Rd., Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 4491)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4491)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2010"; volume: "38"; issue: "6"; pages: "E92"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-DEC-2009) Molecular Biology, Princeton University, Washington 
            Rd., Princeton, NJ 08544, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..426
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    531..671
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(857..1445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1619..2476)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2477..2581)
                     /label=AmpR promoter
     misc_feature    2607..2624
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"
     misc_feature    2625..2649
                     /label=Landing Pad Region 1
                     /note="Landing Pad Region 1"
     primer_bind     2668..2684
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2706..2739
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        2803..3302
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        3314..3361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3380..4378
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRP"
     misc_feature    4415..4448
                     /label=FRT
                     /note="FRT"
     protein_bind    4415..4448
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    4449..4473
                     /label=Landing Pad Region 2
                     /note="Landing Pad Region 2"
     misc_feature    4474..4491
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"
ORIGIN
        1 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
       61 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
      121 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
      181 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
      241 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
      301 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
      361 cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa
      421 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
      481 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
      541 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
      601 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      661 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1921 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2221 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2581 gcacatttcc ccgaaaagtg ccaccttagg gataacaggg taattacggc cccaaggtcc
     2641 aaacggtgaa ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     2701 attccgaagt tcctattctc tagaaagtat aggaacttca ggtctgaaga ggagtttacg
     2761 tccagccaag ctagcttggc tgcaggtcgt cgaaattcta ccgggtaggg gaggcgcttt
     2821 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     2881 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     2941 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     3001 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     3061 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
     3121 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     3181 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     3241 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     3301 cgacctgcag cctgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3361 caaggtgagg aactaaacca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3421 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3481 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3541 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     3601 tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg
     3661 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3721 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg
     3781 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     3841 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     3901 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     3961 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4021 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4081 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4141 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4201 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4261 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4321 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccgg
     4381 atgaggggat caattctcta gagctcgctg atcagaagtt cctattctct agaaagtata
     4441 ggaacttcga tggcgcctca tccctgaagc caatagggat aacagggtaa t
//
