LOCUS       Exported                4945 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  MoClo end-linker vector pELB-1, complete sequence.
ACCESSION   HQ728319
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S.
  TITLE     A modular cloning system for standardized assembly of multigene 
            constructs
  JOURNAL   PLoS ONE 6 (2), E16765 (2011)
  PUBMED    21364738
REFERENCE   2  (bases 1 to 4945)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-DEC-2010) Icon Genetics GmbH, Weinbergweg 22, 
            Halle/Saale 06120, Germany
REFERENCE   3  (bases 1 to 4945)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4945
                     /organism="MoClo end-linker vector pELB-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:993660"
     misc_feature    22..46
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(202..1350)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      1651..2440
                     /allele="ColE1"
     rep_origin      complement(1686..2274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2445..3305)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="ADZ30893.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2445..3305)
                     /gene="bla"
                     /label=bla
     promoter        complement(3306..3410)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3497..4128
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      complement(3503..4120)
                     /direction=LEFT
                     /allele="oriV"
     misc_feature    4220..4244
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4333..4336
                     /note="fusion site; other site"
     protein_bind    4399..4420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4435..4465
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4473..4489
                     /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)."
     primer_bind     4497..4513
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4509..4832
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     gene            4509..4829
                     /gene="lacZ"
                     /label=lacZ
     misc_feature    4526..4582
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4583..4599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4929..4932
                     /note="fusion site; other site"