LOCUS       40924_33942        3293 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning and transformation vector pOT2-3GLA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3293)
  AUTHORS   Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
  TITLE     Plasmid based gap-repair recombineering for Drosophila transgenesis 
            and the analysis of alternative pre-mRNA processing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3293)
  AUTHORS   Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2014) School of Bioscience, University of 
            Birmingham, Edgbaston, Birmingham, West Midlands B15 2TT, United 
            Kingdom
REFERENCE   3  (bases 1 to 3293)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3293)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-OCT-2014) School of Bioscience, University of Birmingham, 
            Edgbaston, Birmingham, West Midlands B15 2TT, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNASTAR Lasergene SeqBuilder v. 7.2.1 
            Sequencing Technology :: not fully sequenced, assembled from pieces,
            vector fully functional
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(188..257)
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     promoter        complement(387..405)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(473..1129)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(1166..1194)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     rep_origin      complement(1256..1844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2004..2095
                     /label=multiple cloning site
                     /note="multiple cloning site"
     protein_bind    2100..2133
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2138..2374
                     /label=3xP3-hsp70
                     /note="3xP3-hsp70"
     regulatory      2369..2378
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2390..3100
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    3137..3236
                     /note="ewg PA1; polyadenylation site 1 of the Drosophila
                     erect wing gene"
     misc_recomb     3248..3281
                     /label=loxP site for cre mediated recombination
                     /note="loxP site for cre mediated recombination"
     protein_bind    complement(3248..3281)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
ORIGIN
        1 tgcccgccgt gaccgtcgag aacccgctga cgctgccccg cgtatccgca cccgccgacg
       61 ccgtcgcacg tcccgtgctc accgtgacca ccgcgcccag cggtttcgag ggcgagggct
      121 tcccggtgcg ccgcgcgttc gccgggatca actaccgcca cctcgacccg ttcatcatga
      181 tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg ccctggcacc
      241 cgcaccgcgg cttcgagacc gtgacctaca tcgtcaccgg cttgacgacc ttggagcgac
      301 tgtcgttagt tccgcgcgat tcggtgcggt atttcacacc gcatatggtg cactctcagt
      361 acaatctgct ctgatgccgc tcgagttcta tagtgtcacc taaatcgtat gtgtatgata
      421 cataaggtta tgtattaatt gtagccgcgt tctaacgtcg acgtaccaat tacgccccgc
      481 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
      541 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
      601 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
      661 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
      721 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
      781 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
      841 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
      901 ttgccatacg gaactccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
      961 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     1021 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     1081 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     1141 tagctcctga aaatctcgat aactcattaa agcttatcga tgataagctg tcaaacatga
     1201 gaatttagcc cgcctaatga gcgggctttt tgttgtgatc cgcgttgctg gcgtttttcc
     1261 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1321 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1381 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1441 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1501 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1561 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1621 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1681 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1741 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1801 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttggatc
     1861 acaaaaaaaa gcccgctcat taggcgggtt aaattcccgg gtactgcagc cgattcatta
     1921 atgcaggtta acctggctta tcgaaattaa tacgactcac tatagggaga ccggcagatc
     1981 tgatatcatc gccactgtgc tgggaattct gtcgacagct gcagtactag tcaagcttag
     2041 gatccgatat ccatggtgct agcgagatct tcctcgaggg taccggcggc cgcgtctaga
     2101 taacttcgta taatgtatgc tatacgaagt tatcatgccg gggatctaat tcaattagag
     2161 actaattcaa ttagagctaa ttcaattagg atccaagctt atcgatttcg aaccctcgac
     2221 cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc aaacaagcaa
     2281 agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga acaagctaaa
     2341 caatcgggcg gccgtactag agccggtcgc caccatggtg tcgagcgcaa gcaagggcga
     2401 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2461 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2521 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2581 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2641 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2701 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2761 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2821 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2881 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2941 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3001 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3061 cgccgccggg atcactctcg gcatggacga gctgtacaag tactcagatc tttgcaagct
     3121 tgtagagttt cccatttaat aattcatatt atctcgaatc tagtcaatta cggctttcct
     3181 caaatagaaa aataaaaaaa atgaaaaaat gcacttgcca tttaaactta gacgcgataa
     3241 gcttgatata acttcgtata atgtatgcta tacgaagtta tacgcgtact aga
//
