LOCUS       Exported                7384 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cosmid vector pPIBTcosTET, complete sequence.
ACCESSION   EU234497
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7384)
  AUTHORS   Oertel W, Weissgerber A.
  TITLE     Transformation of Ulva mutabilis Foyn with plasmids integrating into
            the chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7384)
  AUTHORS   Oertel W, Weissgerber A.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2007) Biochemistry, Genetics and Microbiology, 
            Division of Genetics, University of Regensburg, Universitaetsstr. 
            31, Regensburg, Bavaria 93053, Germany
REFERENCE   3  (bases 1 to 7384)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7384
                     /organism="Cosmid vector pPIBTcosTET"
                     /mol_type="other DNA"
                     /db_xref="taxon:498256"
     misc_feature    1..546
                     /label=contains Ulva mutabilis RbcS promoter and 5 UTR
                     /note="contains Ulva mutabilis RbcS promoter and 5 UTR"
     CDS             join(547..554,1052..1418)
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /note="from Streptoalloteichus hindustanus; adapted to 
                     Chlamydomonas reinhardtii and Ulva mutabilis codon usage"
                     /protein_id="ABZ81849.1"
                     /translation="MAKITSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            547..1418
                     /gene="ble"
                     /label=ble
     intron          555..1051
                     /gene="ble"
                     /note="Ulva mutabilis RbcS intron 1"
     3UTR           1419..1807
                     /label=Ulva mutabilis RbcS 3 UTR
                     /note="Ulva mutabilis RbcS 3 UTR"
     misc_feature    1816..1986
                     /label=cloning vector pUC18, with minor modifications
                     /note="cloning vector pUC18, with minor modifications"
     misc_feature    1968..4852
                     /note="modified EcoRI fragment from pCos2EMBL; contains two
                     cos sites and tetracycline resistance gene"
     CDS             complement(3582..4772)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(4820..4848)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_feature    4855..7384
                     /label=cloning vector pUC18, with minor modifications
                     /note="cloning vector pUC18, with minor modifications"
     CDS             complement(5016..5339)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     5249..5265
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5269..5325
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5335..5351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5359..5375
                     /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(5383..5413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5428..5449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5737..6325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6496..7356)
                     /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"