LOCUS       40924_1994        5311 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector miniTn4001-Puro-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5311)
  AUTHORS   Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.
  TITLE     In situ overlap and sequence synthesis during DNA assembly
  JOURNAL   ACS Synth Biol 2 (12), 750-755 (2013)
  PUBMED    24161008
REFERENCE   2  (bases 1 to 5311)
  AUTHORS   Paetzold B.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2013) EMBL-CRG Systems Biology Research Unit, 
            Centre for Genomic Regulation, Dr. Aiguader 88, Barcelona 08003, 
            Spain
REFERENCE   3  (bases 1 to 5311)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol"; date: "2013"; volume: "2"; issue: "12"; pages: "750-755"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAR-2013) EMBL-CRG Systems Biology Research Unit, Centre for 
            Genomic Regulation, Dr. Aiguader 88, Barcelona 08003, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..18)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  complement(27..52)
                     /mobile_element_type="insertion sequence:IS256"
                     /label=insertion sequence:IS256
     promoter        complement(74..92)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(113..129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(137..153)
                     /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)."
     promoter        complement(161..191)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(206..227)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(515..1103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1277..2134)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2135..2239)
                     /label=AmpR promoter
     rep_origin      complement(2265..2720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2862..2878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2888..2906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3008..4180
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /protein_id="AHL28656.1"
                     /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
                     IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
                     ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
                     LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
                     GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
                     FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
                     ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
     mobile_element  4185..4210
                     /mobile_element_type="insertion sequence:IS256"
                     /label=insertion sequence:IS256
     primer_bind     4222..4238
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4287..4296
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4627..5223
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
ORIGIN
        1 cgatccacta gttctagagc ggccgctttt acacaattat acggacttta tcagctagct
       61 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
      121 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      181 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      241 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      301 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      361 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      421 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      481 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1141 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1201 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1261 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1321 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1381 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1441 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1501 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1561 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1621 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1681 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1741 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1801 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1861 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1921 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1981 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2041 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2101 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2161 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2221 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     2281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2341 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2461 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2641 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2701 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     2761 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2821 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     2881 gcgcgcgtaa tacgactcac tatagggcga attgggtacg aattgtgtaa aagtaaaaag
     2941 gccatataac agtcctttta cggtacaatg tttttaacga caaaaacata cccaggagga
     3001 cttttacatg acccaagtac attttacact gaaaagcgaa gagattcaaa gcattattga
     3061 atattctgta aaggatgacg tttctaaaaa tattttaaca acggtattta atcaactaat
     3121 ggaaaatcaa cgaacagaat atattcaagc aaaagaatat gaacgaacag aaaaccgaca
     3181 aagtcaacga aatggctatt atgagcgcag ctttacgaca cgtgtaggca cgctagaatt
     3241 aaaagtaccc agaacacgtg atggccattt ttcacccaca gtgtttgaac gttatcaacg
     3301 aaacgaaaaa gccctcatgg cttcaatgtt ggaaatgtat gtatcaggcg tttcaactcg
     3361 taaagtatca aaaattgtgg aagaactttg tggtaaatcc gtctctaagt ccttcgtttc
     3421 tagcttaaca gaacagctag aacctatggt taacgagtgg cagaatcgtt tattatcaga
     3481 aaaaaattat ccttacttaa tgaccgatgt actctatata aaagtacgag aagaaaatcg
     3541 agtactctca aaaagctgtc atatagcgat tggaataacc aaagatggcg accgtgaaat
     3601 tatcggcttc atgattcaaa gtggcgaaag cgaagagacc tggacaacat tttttgaata
     3661 cctaaaagaa cgcggtttac aaggtacgga actcgttatt tctgatgcgc acaaaggatt
     3721 agtctctgcc attagaaaat ccttcaccaa cgtaagttgg caaagatgcc aagttcactt
     3781 cctaagaaat atctttacca ccattcctaa aaaaaattca aaatctttca gagaagctgt
     3841 taaaggaatt tttaagttca cagatattaa cttagcgcgt gaggctaaaa atcgattgat
     3901 tcatgattat atcgatcaac caaaatattc aaaagcttgc gcatcattgg atgatggatt
     3961 cgaagacgcc tttcaatata ccgtacaagg aaattcccac aatcgactaa agagtaccaa
     4021 tctaattgaa cgactgaatc aagaagtacg cagaagagaa aagattattc gcatcttccc
     4081 caatcaaaca tcagccaatc gcttaattgg agccgttctt atggacctac atgatgaatg
     4141 gatttattct tcaagaaaat acatcaattt tgataagtag actagataaa gtccgtataa
     4201 ttgtgtaaaa gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct
     4261 gcagcccggg gggcaagatc caatgaccat ggcggccgcc agaattaaaa gttagtgaac
     4321 aagaaaacag tgaagcacca gtttctgaac caaaagaaga cgaaaaaaca aaaaaagatt
     4381 aagcaattta tttggaaaat ctttttttgt ttttttaaga aatatttatt gtttttttta
     4441 aaaaattatg tacaattgct actataaggg aaagaaaaaa agaaagatat aaattgtata
     4501 aagtagggtt agaagcaatt aataattatt attaatgtta tttttctctt atatattcaa
     4561 tgtaatttta attacatttg cttttaataa aaacactact taatagagaa aggaaatata
     4621 agatctatga ctgaatataa acctactgtt agattagcta ctagagatga tgttcctaga
     4681 gctgttagaa ctttagctgc tgcttttgct gattatcctg ctactagaca tactgttgat
     4741 cctgatagac atattgaaag agttactgaa ttacaagaat tatttttaac tagagttggt
     4801 ttagatattg gtaaagtttg ggttgctgat gatggtgctg ctgttgctgt ttggactact
     4861 cctgaaagtg ttgaagctgg tgctgttttt gctgaaattg gtcctagaat ggctgaatta
     4921 agtggtagta gattagctgc tcaacaacaa atggaaggtt tattagctcc tcatagacct
     4981 aaagaacctg cttggttttt agctactgtt ggtgttagtc ctgatcatca aggtaaaggt
     5041 ttaggtagtg ctgttgtttt acctggtgtt gaagctgctg aaagagctgg tgttcctgct
     5101 tttttagaaa ctagtgctcc tagaaattta cctttttatg aaagattagg ttttactgtt
     5161 actgctgatg ttgaagttcc tgaaggtcct agaacttggt gtatgactag aaaacctggt
     5221 gcttaaagat ctaaattaaa gttggtggat cctcgttcaa agttacaaat tggatctttc
     5281 tagaagatct cctacaatat tctcagctgc c
//
