LOCUS       40924_5089        4577 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAV012, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4577)
  AUTHORS   White KP, Victorsen A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2013) Human Genetics, The University of Chicago, 
            5801 South Ellis Avenue, Chicago, IL 60637, USA
REFERENCE   2  (bases 1 to 4577)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4577)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (15-JUL-2013) Human Genetics, The University of Chicago, 5801 South 
            Ellis Avenue, Chicago, IL 60637, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..414
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        509..613
                     /label=AmpR promoter
     CDS             614..1471
                     /label=AmpR
                     /note="beta-lactamase"
     misc_feature    1583..1867
                     /note="sGFP exon 1; 1st exon of superfolder green
                     fluorescent protein"
     primer_bind     1583..1604
                     /label=forward recombineering primer binding site
                     /note="forward recombineering primer binding site"
     misc_feature    1941..1974
                     /label=loxP
                     /note="loxP"
     protein_bind    1941..1974
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        1989..2488
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        2505..2552
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2571..3371
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     protein_bind    complement(3507..3540)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3604..3622
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             4109..4126
                     /label=tetracysteine tag
                     /note="tetracysteine peptide that binds biarsenical
                     labeling reagents"
     CDS             4148..4171
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             4172..4192
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             4235..4258
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     primer_bind     complement(4265..4288)
                     /label=reverse recombineering primer binding site
                     /note="reverse recombineering primer binding site"
     oriT            complement(4334..4443)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta
       61 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat
      121 gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc
      181 tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa
      241 cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta
      301 aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc
      361 cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa
      421 ttagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggatagac
      481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      841 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
      901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1141 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1441 tgagataggt gcctcactga ttaagcattg gtaacccagc ccgcctaatg agcgggcttt
     1501 tttttgaaca aaaggctgca gctggacttc tagagctggt accgaattcg attatgatat
     1561 tccaactact gcaagcgata tcatggtgtc caagggcgag gagctgttca ccggcgtggt
     1621 gcccatcctg gtggagctgg atggcgacgt gaacggccac aagttcagcg tgcgcggcga
     1681 gggcgagggc gacgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1741 gctgcccgtg ccctggccca ccctggtgac caccctgacc tacggcgtgc agtgcttcag
     1801 ccgctacccc gatcacatga agcagcacga tttcttcaag agcgccatgc ccgagggcta
     1861 cgtgcaggta agtgtgtacg gtgggatccg gtctatataa ggagagctgt ctgcaaaccc
     1921 tatgctactg cggccgctcg ataacttcgt atagcataca ttatacgaag ttatggccac
     1981 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     2041 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     2101 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     2161 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     2221 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     2281 aggcctttgg ggcagcggcc aatagcagct ttactccttc gctttctggg ctcagaggct
     2341 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     2401 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     2461 ctcctcttcc tcatctccgg gcctttcgac ctgcagcagc acgtgttgac aattaatcat
     2521 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atgggatcgg
     2581 ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2641 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2701 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2761 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2821 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2881 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2941 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3001 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3061 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccggcg
     3121 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3181 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3241 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3301 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3361 acgagttctt ctgagcggga ctctggggtt cgaataaaga ccgaccaagc gacgtctgag
     3421 agctccctgg cgaattcggt accaataaaa gagctttatt ttcatgatct gtgtgtgttg
     3481 gtttttgtgt gcggcgcgcg aaatgaataa cttcgtatag catacattat acgaagttat
     3541 cgctcgagta ggacaaatcc gccgggataa ttaatacata accttatgta tcatacacat
     3601 acgatttagg tgacactata gaagcttaac atccactttt ttttccccac aggagcgcac
     3661 catcagcttc aaggatgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga
     3721 taccctggtg aaccgcatcg agctgaaggg catcgatttc aaggaggatg gcaacatcct
     3781 gggccacaag ctggagtaca acttcaacag ccacaacgtg tacatcaccg ccgataagca
     3841 gaagaacggc atcaaggcca acttcaagat ccgccacaat gtggaggatg gctccgtgca
     3901 gctggccgat cactaccagc agaacacccc catcggcgac ggcccagtgc tgctgcccga
     3961 taaccactac ctgagcaccc agagcgtgct gtccaaggac cccaacgaga agcgcgatca
     4021 catggtgctg ctggagttcg tgaccgccgc cggcatcacc ctgggcatgg atgagctgta
     4081 caaggagagc agcggcagct tcctgaactg ctgcccagga tgctgcatgg aaccgggaag
     4141 cggcaactgg tcccaccccc agttcgagaa ggaaaacctg tacttccaga gcgattacaa
     4201 ggatcacgac ggcgactaca aggaccacga tattgattac aaggacgatg acgacaagag
     4261 cgctctgacc gacgacctgc ccagggcctt ccggtcgacc tcgagttgcc ctcatctgtt
     4321 acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc
     4381 gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg
     4441 cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct
     4501 gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag
     4561 ggttatgcag cggaaaa
//
