LOCUS       Exported                4968 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1F-3, complete sequence.
ACCESSION   KF955552
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  AUTHORS   Kong DS, Thorsen TA, Babb J, Wick S, Gam JJ, Weiss R, Carr PA.
  TITLE     Microfluidic Assembly of Genetic Circuits
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4968)
  AUTHORS   Gam JJ, Bram E, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-DEC-2013) Biological Engineering, MIT, 77 Mass Ave 
            (NE47-215), Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4968)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4968
                     /organism="Cloning vector pL1F-3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1469142"
     protein_bind    96..117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        132..162
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    170..186
                     /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     194..210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             206..529
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="AHK23083.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     gene            206..529
                     /gene="lacZ"
                     /label=lacZ
     misc_feature    223..279
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(280..296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    626..629
                     /label=fusion site
                     /note="fusion site"
     misc_feature    630..633
                     /label=fusion site
                     /note="fusion site"
     misc_feature    671..695
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(851..1999)
                     /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      2300..3089
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3094..3954)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="AHK23082.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3094..3954)
                     /gene="bla"
                     /label=bla
     promoter        complement(3955..4059)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4146..4777
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      complement(4152..4769)
                     /direction=LEFT
                     /label=oriV
                     /note="oriV"
     misc_feature    4869..4893
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4936..4939
                     /label=fusion site
                     /note="fusion site"
     misc_feature    4952..4955
                     /label=fusion site
                     /note="fusion site"