LOCUS       Exported                3965 bp DNA     circular SYN 05-JUL-2019
DEFINITION  Standard E. coli vector with a multiple cloning site (MCS) for DNA 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    #3 pEF1alpha-rTetR-bGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3965)
  AUTHORS   Yanisch-Perron C, Vieira J, Messing J.
  TITLE     Improved M13 phage cloning vectors and host strains: nucleotide 
            sequences of the M13mp18 and pUC19 vectors.
  JOURNAL   Gene 1985;33:103-19.
  PUBMED    2985470
REFERENCE   2  (bases 1 to 3965)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 4, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1985"; volume: "33"; pages: "103-19"
COMMENT     See also GenBank accession L09137.
FEATURES             Location/Qualifiers
     source          1..3965
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        2..213
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          214..1152
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1109..1129
                     /label=EF1alpha_seq_Fwd
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1144..1167)
                     /label=EF1alpha_BamHI_Rv
     primer_bind     1161..1183
                     /label=rTetR_BamHI_Fwd
     CDS             1171..1917
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(1899..1923)
                     /label=rTetR_HindIII_Rv
     primer_bind     1916..1944
                     /label=bGHpA_Fwd_HindIII
     misc_feature    1918..1923
                     /label=MCS
                     /note="pUC19 multiple cloning site"
     polyA_signal    1924..2148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2130..2154)
                     /label=bGHpA_Rv_PciI
     rep_origin      complement(2210..2798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2969..3829)
                     /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"
     promoter        complement(3830..3934)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3960..3965,1..18)
                     /label=EF1alpha_Fwd_AatII
ORIGIN
        1 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
       61 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
      121 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
      181 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
      241 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
      301 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
      361 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
      421 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
      481 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
      541 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
      601 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
      661 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
      721 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
      781 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
      841 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
      901 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
      961 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     1021 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     1081 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     1141 cttccatttc aggtgtcgtg aggatccacc atgtctagac tggacaagag caaagtcata
     1201 aacggcgctc tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc
     1261 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     1321 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg
     1381 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc
     1441 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     1501 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     1561 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     1621 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     1681 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     1741 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac
     1801 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     1861 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaaag
     1921 cttctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1981 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2041 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2101 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggac atgtgagcaa
     2161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3961 acgtc
//
