LOCUS       40924_1984        5321 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector Mini-Tn4001PSpuro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  AUTHORS   Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman
            C.
  TITLE     New selectable marker for manipulating the simple genomes of 
            Mycoplasma species
  JOURNAL   Antimicrob. Agents Chemother. 53 (10), 4429-4432 (2009)
  PUBMED    19687239
REFERENCE   2  (bases 1 to 5321)
  AUTHORS   Merryman C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2009) Synthetic Biology, J. Craig Venter 
            Institute, 9704 Medical Center Dr, Rockville, MD 20850, USA
REFERENCE   3  (bases 1 to 5321)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5321)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
            Agents Chemother."; date: "2009"; volume: "53"; issue: "10"; pages: 
            "4429-4432"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2009) Synthetic Biology, J. Craig Venter Institute, 9704 
            Medical Center Dr, Rockville, MD 20850, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..600)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             1105..2277
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /protein_id="ACR82773.1"
                     /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
                     IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
                     ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
                     LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
                     GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
                     FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
                     ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
     gene            1105..2277
                     /gene="tnp"
                     /label=tnp
     promoter        complement(2298..2316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2323..2339)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2481..2936
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2962..3066
                     /label=AmpR promoter
     CDS             3067..3924
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4098..4686
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4974..4995
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5010..5040
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5064
                     /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     5072..5088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5109..5127
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaagcacca ggttttctag tcatacacca agttctagga ccttcaggaa cttcaacatc
       61 agcagtaaca gtaaaaccta atctttcata aaaaggtaaa tttctaggag cactagtttc
      121 taaaaaagca ggaacaccag ctctttcagc agcttcaaca ccaggtaaaa caacagcact
      181 acctaaacct ttaccttgat gatcaggact aacaccaaca gtagctaaaa accaagcagg
      241 ttctttaggt ctatgaggag ctaataaacc ttccatttgt tgttgagcag ctaatctact
      301 accacttaat tcagccattc taggaccaat ttcagcaaaa acagcaccag cttcaacact
      361 ttcaggagta gtccaaacag caacagcagc accatcatca gcaacccaaa ctttaccaat
      421 atctaaacca actctagtta aaaataattc ttgtaattca gtaactcttt caatatgtct
      481 atcaggatca acagtatgtc tagtagcagg ataatcagca aaagcagcag ctaaagttct
      541 aacagctcta ggaacatcat ctctagtagc taatctaaca gtaggtttat attcagtcat
      601 agatcttata tttcctttct ctattaagta gtgtttttat taaaagcaaa tgtaattaaa
      661 attacattga atatataaga gaaaaataac attaataata attattaatt gcttctaacc
      721 ctactttata caatttatat ctttcttttt ttctttccct tatagtagca attgtacata
      781 attttttaaa aaaaacaata aatatttctt aaaaaaacaa aaaaagattt tccaaataaa
      841 ttgcttaatc tttttttgtt ttttcgtctt cttttggttc agaaactggt gcttcactgt
      901 tttcttgttc actaactttt aattctgcag gtcgactcta gaggatcgga tccccgggta
      961 ccgagctcga attcttttac acaattatac ggactttatc aagcttatcg ataccgtcga
     1021 cgtgtaaaag taaaaaggcc atataacagt ccttttacgg tacaatgttt ttaacgacaa
     1081 aaacataccc aggaggactt ttacatgacc caagtacatt ttacactgaa aagcgaagag
     1141 attcaaagca ttattgaata ttctgtaaag gatgacgttt ctaaaaatat tttaacaacg
     1201 gtatttaatc aactaatgga aaatcaacga acagaatata ttcaagcaaa agaatatgaa
     1261 cgaacagaaa accgacaaag tcaacgaaat ggctattatg agcgcagctt tacgacacgt
     1321 gtaggcacgc tagaattaaa agtacccaga acacgtgatg gccatttttc acccacagtg
     1381 tttgaacgtt atcaacgaaa cgaaaaagcc ctcatggctt caatgttgga aatgtatgta
     1441 tcaggcgttt caactcgtaa agtatcaaaa attgtggaag aactttgtgg taaatccgtc
     1501 tctaagtcct tcgtttctag cttaacagaa cagctagaac ctatggttaa cgagtggcag
     1561 aatcgtttat tatcagaaaa aaattatcct tacttaatga ccgatgtact ctatataaaa
     1621 gtacgagaag aaaatcgagt actctcaaaa agctgtcata tagcgattgg aataaccaaa
     1681 gatggcgacc gtgaaattat cggcttcatg attcaaagtg gcgaaagcga agagacctgg
     1741 acaacatttt ttgaatacct aaaagaacgc ggtttacaag gtacggaact cgttatttct
     1801 gatgcgcaca aaggattagt ctctgccatt agaaaatcct tcaccaacgt aagttggcaa
     1861 agatgccaag ttcacttcct aagaaatatc tttaccacca ttcctaaaaa aaattcaaaa
     1921 tctttcagag aagctgttaa aggaattttt aagttcacag atattaactt agcgcgtgag
     1981 gctaaaaatc gattgattca tgattatatc gatcaaccaa aatattcaaa agcttgcgca
     2041 tcattggatg atggattcga agacgccttt caatataccg tacaaggaaa ttcccacaat
     2101 cgactaaaga gtaccaatct aattgaacga ctgaatcaag aagtacgcag aagagaaaag
     2161 attattcgca tcttccccaa tcaaacatca gccaatcgct taattggagc cgttcttatg
     2221 gacctacatg atgaatggat ttattcttca agaaaataca tcaattttga taagtagaaa
     2281 tggggtaccc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     2341 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2401 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2461 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2581 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2641 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2701 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2761 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2821 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2881 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     2941 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3001 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3061 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3121 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3181 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3241 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3301 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3361 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3421 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3481 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3541 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3601 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3661 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3721 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3781 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3841 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3901 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3961 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4021 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4681 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4861 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4921 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4981 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5041 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     5101 aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg agctcagtca agtccagact
     5161 cctgtgtaaa aactagtgga tccgcttgca tgcctgcagt tcagatcctt taaaaaataa
     5221 aaaacaacct tttggttgtt ttttaataaa ttaattaata attaattatt cactttttaa
     5281 acgaattgta actttgaatg aaccaacttt aatttagatc t
//
