LOCUS       40924_5884        5023 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAM1-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5023)
  AUTHORS   Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
  TITLE     pBAM1: an all-synthetic genetic tool for analysis and construction 
            of complex bacterial phenotypes
  JOURNAL   BMC Microbiol. 11 (1), 38 (2011)
  PUBMED    21342504
REFERENCE   2  (bases 1 to 5023)
  AUTHORS   Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2011) Systems Biology Program, Centro Nacional de 
            Biotecnologia, C/ Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 5023)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5023)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Microbiol."; date: "2011"; volume: "11"; issue: "1"; pages: "38"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JAN-2011) Systems Biology Program, Centro Nacional de 
            Biotecnologia, C/ Darwin 3, Madrid, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(12..400)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            complement(419..527)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(674..713)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(720..1577)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1578..1669)
                     /label=AmpR promoter
     CDS             complement(1726..3153)
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     misc_feature    3247..3265
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             3274..3984
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4102..4914
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    complement(5002..5020)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 ggcgcgcccc atgtcagccg ttaagtgttc ctgtgtcact caaaattgct ttgagaggct
       61 ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa
      121 ggctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat
      181 ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca
      241 tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt
      301 taaacatgag agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta
      361 gtacgtacta tcaacaggtt gaactgctga tcttcagatc ggcgcgccgg ccggcctacg
      421 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
      481 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
      541 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
      601 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
      661 aggacaaggg tcgtccaaaa aaaaaggctc caaaaggagc ctttaattgt atcggtttac
      721 caatgtttaa tcaggctcgc gccaatttcc gcaatctgac ggttacgttc atccatcgtc
      781 gcctggctgc cggtggtata aatcaccaca atacggctcg gtttaccatc cgggcccagc
      841 gccgcaataa tgccacggct gccacgttcg cccgcaccgc ttttatccgc aataaaccag
      901 ccagccggca gcgcgctacg cagcagcgga cccgccactt tatccgcttc catccaatca
      961 atcagttgct ggcggcttgc cagggtcagc agctcacccg tcagcagttt acgcagggtg
     1021 gtcgccattg ctgccggcat ggtggtatca cgttcatcgt tcggaatcgc ttcgttcagt
     1081 tccggttccc aacgatccag acgggtcaca tgatcgccca tgttatgcag aaacgcggtc
     1141 agttctttcg gaccgccaat ggtcgtcagc agcaggttcg ccgcggtgtt atcgctcatg
     1201 gtaatcgccg cgctgcacag ttcacgcacg gtcatgccat cggtcagatg tttttcggtc
     1261 accgggctat attccaccag atcgttctgg ctataatgaa tacgacggcc cagttgttcc
     1321 tggcccgcat ccacacggct cagaaccgca ccgcacagca gcactttaaa ggtgctcatc
     1381 atcggaaaac gttcttccgg acgaaaagat tccagaattt tgccgctgtt cagatccagt
     1441 tcaatatagc ccacgcgcgc acccagttga tcttccgcat ctttcacttt caccagggtt
     1501 tccggatgcg caaacaccgg caggcaaaac gccgcaaaaa acggaatcag cgccacacga
     1561 aaatgctgaa tgctcatact cttccttttt caatattatt gaagcattta tcagggttat
     1621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggaatta
     1681 aaaaaaagcc cgctcattag gcgggctact aatttaaatc gattagattt taatgccctg
     1741 cgccatcagg tctttcgcgg ccagaaagcc atccagtttg ctttgcagcg cttcccaacc
     1801 ttcccacagc gcaccccagc tcgcaatgcc ggtacgtttg ctatccataa agccgcccag
     1861 acgcgcaatc gccatatacg cccattgcag gctgcccgct ttttctttgc gtttgcgttt
     1921 gcctttatcc agatagccca gcagttggca ttcatccggg gtcagcacgg tttccgcgct
     1981 ctggctttca acgtgttccg cttctttcag caggccctgc gcacgcagtg cttgcggcgg
     2041 agtaaaagat tcacgcagtt gcagcagacg caccgccaca aagctcagaa tgctcaccat
     2101 acgttccagg ttatccggtt cttccatacg ctgacgttcc gcacccgcac ccgttttcca
     2161 cgctttgtga aattcttcaa tgcgccaacg atgggtataa atatcaatca cacgcagcgc
     2221 ttgggccaga ctttccaccg gctcgctggt cagcagcagc catttcagcg gggtttcgcc
     2281 tttcggcgga ttaatttctt cggccagcac cgcgttcagg gtaatgttgc cctgtttcag
     2341 ggtaatacgg ccgctacgca ggctcaggct cgctttacgc gccggacggt ttttacgttt
     2401 gccacgttta tccaccacgc ctttctgcgg aatgctaatc tgatagccgc ccagttccgg
     2461 ctggtttttc aggtgatcat acagatacag gccgctttcc acatctttac gcggatgttt
     2521 gctacgcacc acaaaacgtt cgttatgggc cagtttatct tgcagatacg catgaatatc
     2581 cgcttcacga tcgcacaccg caatcacgtt gctcatcatg ctgcccattc tcagacgcga
     2641 agttgcagca gcggccagcc atttgccgct ttctttttca tccgcatccg ccggatcatc
     2701 cggacgcatc caccattctt gatgcagcag gcccacggta cgaaaggtgg tcgcttccag
     2761 cagcagcacg ctatgcaccc accaaccacg gcttttatcc tgaatgctac ccagtttgcc
     2821 cagttcttcc gccacctgat gacgatagct cagagaggtg gtatcttcaa ttgccagcag
     2881 ttccggaaat tcctgggcca gtttcacggt ctgcatggca cccgctttac gaatcgcttc
     2941 cgcgctcacg ttcggattac gaataaaacg atacgcgcct tcctgcatgg ctttgctgcc
     3001 ttcgctgcta atggtaatgc ttttgccgct atatttggcc agttgcgccg caacattcac
     3061 cagacgcgcg gtacgacgcg gatcacccag cgcagcacta gaaaacacgc ttttcgccca
     3121 atccgccgca cgatgcagtg cactggtaat catgaacgtt accatgttag gaggtcacat
     3181 ggaacatcag attctggaaa acgggaaagg ttccgttcag gacgctactt gtgtagttta
     3241 aaccagctgt ctcttataca catctcctgc aggagtaaag gagaagaact tttcactgga
     3301 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     3361 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     3421 ggaaaactac ctgttccatg gccaacactt gtcactactt tgacttatgg tgttcaatgc
     3481 ttttcaagat acccagatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
     3541 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
     3601 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     3661 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     3721 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
     3781 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
     3841 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     3901 cccaacgaaa agagagacca catggtcctt cttgagtttg taaccgctgc tgggattaca
     3961 catggcatgg atgaactata caaagcggcc gcatttaaat ttgtgtctca aaatctctga
     4021 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     4081 aacagtaata caaggggtgt tatgagccat attcagcgtg aaacgagctg tagccgtccg
     4141 cgtctgaaca gcaacatgga tgcggatctg tatggctata aatgggcgcg tgataacgtg
     4201 ggtcagagcg gcgcgaccat ttatcgtctg tatggcaaac cggatgcgcc ggaactgttt
     4261 ctgaaacatg gcaaaggcag cgtggcgaac gatgtgaccg atgaaatggt gcgtctgaac
     4321 tggctgaccg aatttatgcc gctgccgacc attaaacatt ttattcgcac cccggatgat
     4381 gcgtggctgc tgaccaccgc gattccgggc aaaaccgcgt ttcaggtgct ggaagaatat
     4441 ccggatagcg gcgaaaacat tgtggatgcg ctggccgtgt ttctgcgtcg tctgcatagc
     4501 attccggtgt gcaactgccc gtttaacagc gatcgtgtgt ttcgtctggc ccaggcgcag
     4561 agccgtatga acaacggcct ggtggatgcg agcgattttg atgatgaacg taacggctgg
     4621 ccggtggaac aggtgtggaa agaaatgcat aaactgctgc cgtttagccc ggatagcgtg
     4681 gtgacccacg gcgattttag cctggataac ctgattttcg atgaaggcaa actgattggc
     4741 tgcattgatg tgggccgtgt gggcattgcg gatcgttatc aggatctggc cattctgtgg
     4801 aactgcctgg gcgaatttag cccgagcctg caaaaacgtc tgtttcagaa atatggcatt
     4861 gataatccgg atatgaacaa actgcaattt catctgatgc tggatgaatt tttctaataa
     4921 ttaattggac cgcggtccgc gcgggtcctg caggggtgaa agcaacgtgg tggtgcatca
     4981 ggcggatgaa cgcgcggccg cagatgtgta taagagacag ctg
//
