LOCUS       pminiTol2                3459 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pminiTol2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3459)
  AUTHORS   Balciunas D, Wangensteen KJ, Wilber A, Bell J, Geurts A, Sivasubbu 
            S, Wang X, Hackett PB, Largaespada DA, McIvor RS, Ekker SC.
  TITLE     Harnessing a high cargo-capacity transposon for genetic applications
            in vertebrates.
  JOURNAL   PLoS Genet. 2006 Nov 10;2(11):e169. Epub 2006 Aug 28.
COMMENT     This file is created by Vector NTI
            //gt12365.cn/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|gt12365.cn|
COMMENT     VNTNAME|pminiTol2|
FEATURES             Location/Qualifiers
     source          1..3459
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(584..602)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(585..602)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(620..636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(620..636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(633..655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    644..660
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(668..698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    713..734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(851..868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1022..1610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1102..1121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1781..2641)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2404..2423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2642..2746)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2824..3279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2961..2982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3173..3192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3405..3427
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3419..3436
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3420..3436
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3443..3
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3443..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccagagg tgtaaagtac ttgagtaatt ttacttgatt actgtactta
       61 agtattattt ttggggattt ttactttact tgagtacaat taaaaatcaa tacttttact
      121 tttacttaat tacatttttt tagaaaaaaa agtacttttt actccttaca attttattta
      181 cagtcaaaaa gtacttattt tttggagatc acttcattct attttccctt gctattacca
      241 aaccaattga attgcgctga tgcccagttt aatttaaata gatctggcca tctagagcgg
      301 ccgcgcgcac tagtgaattc catggatatc aagcttaaac aagaatctct agttttcttt
      361 cttgctttta cttttacttc cttaatactc aagtacaatt ttaatggagt acttttttac
      421 ttttactcaa gtaagattct agccagatac ttttactttt aattgagtaa aattttccct
      481 aagtacttgt actttcactt gagtaaaatt tttgagtact ttttacacct ctgctcgacc
      541 atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag tgtcacctaa
      601 atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      661 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      721 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      781 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1441 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2641 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2701 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2761 aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2821 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2881 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2941 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3001 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3061 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3121 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3181 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3241 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     3301 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     3361 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     3421 taaaacgacg gccagtgaat tgtaatacga ctcactata
//