LOCUS       Exported                5510 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pPICBT2, complete sequence.
ACCESSION   EU194350
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  AUTHORS   Oertel W, Weissgerber A.
  TITLE     Transformation of Ulva mutabilis Foyn with plasmids integrating into
            the chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5510)
  AUTHORS   Oertel W, Weissgerber A.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-2007) Biochemistry, Genetics and Microbiology, 
            Division of Genetics, University of Regensburg, Universitaetsstr. 
            31, Regensburg, Bavaria 93053, Germany
REFERENCE   3  (bases 1 to 5510)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5510
                     /organism="Transformation vector pPICBT2"
                     /lab_host="Ulva mutabilis"
                     /mol_type="other DNA"
                     /db_xref="taxon:485102"
     regulatory      1..546
                     /regulatory_class="promoter"
                     /note="Ulva mutabilis RbcS promoter"
     CDS             join(547..554,2051..2417)
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /note="derived from Streptoalloteichus hindustanus; adapted
                     to Chlamydomonas reinhardtii and Ulva mutabilis codon 
                     usage"
                     /protein_id="ABX56856.1"
                     /translation="MAKITSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            547..2417
                     /gene="ble"
                     /label=ble
     intron          join(555..717,1718..2050)
                     /gene="ble"
                     /note="Ulva mutabilis RbcS intron 1"
     CDS             complement(862..1521)
                     /codon_start=1
                     /gene="CAT"
                     /product="chloramphenicol acetyl transferase"
                     /label=CAT
                     /note="Cm resistance gene from pBR328"
                     /protein_id="ABX56857.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(862..1521)
                     /gene="CAT"
                     /label=CAT
                     /note="Cm"
     promoter        complement(1522..1624)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     regulatory      2418..2806
                     /regulatory_class="terminator"
                     /note="Ulva mutabilis RbcS terminator"
     misc_feature    2807..5510
                     /label=cloning vector pUC18 with minor modifications
                     /note="cloning vector pUC18 with minor modifications"
     CDS             complement(3142..3465)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     3375..3391
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3395..3451
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3461..3477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3485..3501
                     /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(3509..3539)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3554..3575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3863..4451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4622..5482)
                     /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"