LOCUS       Exported                4010 bp ds-DNA     circular SYN 15-MAY-2019
DEFINITION  Complementary copy of pBR322_NEB.
ACCESSION   .
VERSION     .
KEYWORDS    pBR322_KANR_AgeI_AvrII_PSE1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4010)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 15, 2019 from SnapGene 4.1.3
            http://www.snapgene.com
COMMENT     1.  pBR322 beta-lactamase (bla, conferring ampicillin resistance) is
            transcribed in vitro and in vivo both from its natural promoter (P3)
            and from promoter P1, a pSC101 promoter artificially juxtaposed with
            the bla sequence during construction of the pBR322 plasmid.  (Ref: 
            Brosius et al, J Biol Chem 257, 9205-9210 (1982)). 2.  Corrects 
            sequence error in tet gene deriving from the original pSC101 
            sequence, present in GenBank entry (see accession J01749) and NEB 
            sequences dated 6/6/02 and earlier, based on sequencing of pBR322 
            and pACYC184 at NEB in 2007 (Mathieu-Sheltry, V. and Cantin, E., 
            unpublished).  Correction is C1134T, representing a Thr350Ile 
            correction in the expressed protein.
            ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..4010
                     /organism="Cloning vector pBR322"
                     /mol_type="other DNA"
     source          271..1101
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          3110..3925
                     /organism="Cloning vector pBR322"
                     /mol_type="other DNA"
     -35_signal      139..144
                     /label=-35_Signal _5
                     /note="bla (P3) promoter (counter-clockwise); TTCAAA"
     -10_signal      160..165
                     /label=-10_Signal _5
                     /note="bla (P3) promoter (counter-clockwise); GAGACA; 
                     transcript start 4188 (complementary strand)"
     RBS             197..201
                     /label=RBS _2
                     /note="bla RBS"
     sig_peptide     209..277
                     /gene="bla"
                     /note="bla required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     gene            209..270
                     /gene="bla"
                     /label=bla
                     /note="bla"
     CDS             278..1090
                     /codon_start=1
                     /label=PSE1
                     /note="PSE1"
                     /translation="SSSKFQQVEQDVKAIEVSLSARIGVSVLDTQNGEYWDYNGNQRFP
                     LTSTFKTIACAKLLYDAEQGKVNPNSTVEIKKADLVTYSPVIEKQVGQAITLDDACFAT
                     MTTSDNTAANIILSAVGGPKGVTDFLRQIGDKETRLDRIEPDLNEGKLGDLRDTTTPKA
                     IASTLNKFLFGSALSEMNQKKLESWMVNNQVTGNLLRSVLPAGWNIADRSGAGGFGARS
                     ITAVVWSEHQAPIIVSIYLAQTQASMAERNDAIVKIGHSIFDVYTSQSR"
     rep_origin      1264..1852
                     /label=Rep_Origin _1
                     /note="pMB1 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     CDS             complement(2280..2471)
                     /codon_start=1
                     /exception="alternative start codon"
                     /gene="rop"
                     /product="ROP protein"
                     /label=rop
                     /note="rop"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(2280..2471)
                     /gene="rop"
                     /label=rop
                     /note="rop"
     CDS             complement(3110..3925)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein, class C"
                     /label=KanR
                     /note="Kan tetR (confers resistance to tetracycline)"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(3110..3925)
                     /gene="tet"
                     /label=tet
                     /note="tet"
     -35_signal      3936..3947
                     /label=-35_Signal _2
                     /note="P1 promoter -35 sequence region; transcript starts 
                     36, 37 (complementary strand)"
     -10_signal      3962..3967
                     /label=-10_Signal _2
                     /note="P1 promoter (counter-clockwise); TAAACT"
     -10_signal      complement(3973..3978)
                     /label=-10_Signal _1
                     /note="tet (P2) promoter (clockwise); TTTAAT; transcript 
                     start 45"
     -35_signal      complement(3996..4001)
                     /label=-35_Signal _1
                     /note="tet (P2) promoter (clockwise); TTGACA"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gcctaccggt ttttgctagt agttcaaagt ttcagcaagt
      301 tgaacaagac gttaaggcaa ttgaagtttc tctttctgct cgtataggtg tttccgttct
      361 tgatactcaa aatggagaat attgggatta caatggcaat cagcgcttcc cgttaacaag
      421 tacttttaaa acaatagctt gcgctaaatt actatatgat gctgagcaag gaaaagttaa
      481 tcccaatagt acagtcgaga ttaagaaagc agatcttgtg acctattccc ctgtaataga
      541 aaagcaagta gggcaggcaa tcacactcga tgatgcgtgc ttcgcaacta tgactacaag
      601 tgataatact gcggcaaata tcatcctaag tgctgtaggt ggccccaaag gcgttactga
      661 ttttttaaga caaattgggg acaaagagac tcgtctagac cgtattgagc ctgatttaaa
      721 tgaaggtaag ctcggtgatt tgagggatac gacaactcct aaggcaatag ccagtacttt
      781 gaataaattt ttatttggtt ccgcgctatc tgaaatgaac cagaaaaaat tagagtcttg
      841 gatggtgaac aatcaagtca ctggtaattt actacgttca gtattgccgg cgggatggaa
      901 cattgcggat cgctcaggtg ctggcggatt tggtgctcgg agtattacag cagttgtgtg
      961 gagtgagcat caagccccaa ttattgtgag catctatcta gctcaaacac aggcttcaat
     1021 ggcagagcga aatgatgcga ttgttaaaat tggtcattca atttttgacg tttatacatc
     1081 acagtcgcgc taactgtcag acctaggtta ctcatatata ctttagattg atttaaaact
     1141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1381 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2101 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     2161 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2281 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     2341 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     2401 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     2461 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     2521 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     2581 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     2641 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     2701 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     2761 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     2821 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     2881 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     2941 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     3001 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     3061 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt cagaaaaact
     3121 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     3181 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     3241 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     3301 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     3361 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     3421 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3481 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3541 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3601 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     3661 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     3721 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     3781 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3841 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     3901 acgtttcccg ttgaatatgg ctcatacacg gtgcctgact gcgttagcaa tttaactgtg
     3961 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa
//
