LOCUS       40924_36293        6103 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR6KT-miniTn7T-P1eGFP-FK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6103)
  AUTHORS   Kvitko B, Bruckbauer S.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-FEB-2013) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   2  (bases 1 to 6103)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6103)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (05-FEB-2013) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    18..65
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(175..966)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1352..1399
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1562..2278)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICATGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFKGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(2604..2632)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     mobile_element  complement(2708..2873)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3177..3285
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3308..3692
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(4104..4961)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4962..5066)
                     /label=AmpR promoter
     repeat_region   5543..5741
                     /label=Tn7R
                     /note="Tn7R"
     primer_bind     5746..5762
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5790..5884
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      5987..6073
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ctcgaattgg ggatcttgaa gtacctattc cgaagttcct attctctaga aagtatagga
       61 acttcagagc gcttttgaag ctgatgtgct taaaaactta ctcaatggaa taattctaga
      121 taattcacta gtgatggaaa tgttggatcc tcatactctt cctttttcaa ttcagaagaa
      181 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
      241 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
      301 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
      361 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
      421 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
      481 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
      541 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
      601 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gggatgacag
      661 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
      721 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
      781 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
      841 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
      901 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
      961 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     1021 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     1081 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta
     1141 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt
     1201 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     1261 ctacgtgttc cgcttccttt agcagccctt gcgccatgca tttgttaatc gaattatcca
     1321 ttgctgttga caaagggaat caggggatct tgaagttcct attccgaagt tcctattctc
     1381 tagaaagtat aggaacttca gagcgctttt gaagctaatt cgagatcatg catgagctca
     1441 ctagtggatc ccccgggctg caggaattcg gcttactagt ttagatgcgg tggatgccgg
     1501 ggccgcggta gcgccgggag cagaactcga aggaggagct ggagatctgg gtccggccgc
     1561 tcttgtacag ctcgtccatg cccagggtga tgccggcggc ggtcacgaac tccagcagga
     1621 ccatgtggtc ccgcttctcg ttcgggtcct tgctcagggc gctctgggtg gacaggtagt
     1681 ggttgtccgg cagcaggacc gggccgtcgc cgatcggggt gttctgctgg tagtggtccg
     1741 ccagctggac cgagccgtcc tcgatgttgt ggcggatctt gaagttgacc ttgatgccgt
     1801 tcttctgctt gtcggccatg atgtagacgt tgtgggagtt gtagttgtac tccagcttgt
     1861 ggcccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttgacca
     1921 gggtgtcgcc cttgaacttc acctccgcgc gggtcttgta gttgccgtcg tccttgaaga
     1981 agatggtgcg ctcctggacg tagccctccg gcatcgccga cttgaagaag tcgtgctgct
     2041 tcatgtggtc cgggtagcgc gagaagcact ggacgccgta ggtcagggtc gtgaccaggg
     2101 tgggccacgg gacgggcagc ttgccggtgg cgcagatgaa cttcagggtc agcttgccgt
     2161 aggtggcgtc gccctcgccc tcgcccgaca ccgagaactt gtggccgttg acgtcgccgt
     2221 ccagctcgac caggatgggc acgacgccgg tgaacagctc ctcgcccttg ctcaccatgt
     2281 cgaatccttc ttgtgaatct attatggcga caagcaaatc gagctctgac tgcctacccc
     2341 acaacaacta tcagaaagca ccagcacaac ggctgcctaa ctttgtttta gggcgactgc
     2401 cctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     2461 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     2521 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     2581 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     2641 gtgttcacta gtaagccgaa ttcctcgaga agcttgggcc cggtacctcg cgaaggcctt
     2701 gcaggccaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     2761 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     2821 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     2881 atttttcttc ctgttatgtt tgggcgctgc attaatgaat cggccaacgc gcggggagag
     2941 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgacccgctg cgctcggtcg
     3001 ttcggctgcg gcgagcggta tcaggggatt ccttaaggta tactttccgc tgcataaccc
     3061 tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca
     3121 ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagccgggc
     3181 aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt cccttattcg
     3241 cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgataag ctctgataac
     3301 gcaggaaaga acatgggcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca
     3361 cgtactaagc tctcaagttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat
     3421 ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc
     3481 tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt
     3541 aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt
     3601 gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt
     3661 ttgagtgaca caggaacact taacggctga caaccggctg gtaacaggat tagcagagcg
     3721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3781 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5101 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5161 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5221 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5281 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5341 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     5401 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5461 ttacgccagc tagaggacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa
     5521 atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag
     5581 atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc
     5641 cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt
     5701 tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gggttcgagg tcgacggtat
     5761 cgataagcta gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc
     5821 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     5881 gaatccaagc tagactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     5941 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     6001 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     6061 gagtaggaca aatccgccgc taggagcttg cggcccggac gat
//
