LOCUS       Exported               13407 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTARBAC6, complete sequence.
ACCESSION   EF081457
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13407)
  AUTHORS   Osoegawa K, Vessere GM, Li Shu C, Hoskins RA, Abad JP, de Pablos B, 
            Villasante A, de Jong PJ.
  TITLE     BAC clones generated from sheared DNA
  JOURNAL   Genomics 89 (2), 291-299 (2007)
  PUBMED    17098394
REFERENCE   2  (bases 1 to 13407)
  AUTHORS   Osoegawa K, De Jong PJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-OCT-2006) Childrens Hospital and Research Center at 
            Oakland, 747 52nd Street, Oakland, CA 94609, USA
REFERENCE   3  (bases 1 to 13407)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13407
                     /organism="Cloning vector pTARBAC6"
                     /mol_type="other DNA"
                     /db_xref="taxon:412753"
     primer_bind     complement(115..131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    139..155
                     /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(163..193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    208..229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(517..1105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1276..2136)
                     /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"
     promoter        complement(2137..2241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2715..2731
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2757..2775)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             2875..4293
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="ABK79083.1"
                     /translation="MNIKKIVKQATVLTFTTALLAGGATQAFAKENNQKAYKETYGVSH
                     ITRHDMLQIPKQQQNEKYQVPQFDQSTIKNIESAKGLDVWDSWPLQNADGTVAEYNGYH
                     VVFALAGSPKDADDTSIYMFYQKVGDNSIDSWKNAGRVFKDSDKFDANDPILKDQTQEW
                     SGSATFTSDGKIRLFYTDYSGKHYGKQSLTTAQVNVSKSDDTLKINGVEDHKTIFDGDG
                     KTYQNVQQFIDEGNYTSGDNHTLRDPHYVEDKGHKYLVFEANTGTENGYQGEESLFNKA
                     YYGGGTNFFRKESQKLQQSAKKRDAELANGALGIIELNNDYTLKKVMKPLITSNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGINSNDIYMLGYVSNSLTGPYKPLNKTGLVLQ
                     MGLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFFEDKKATFAPSFLMNIKGNKTSV
                     VKNSILEQGQLTVN"
     gene            2875..4293
                     /gene="sacB"
                     /label=sacB
     promoter        5243..5430
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             5431..6093
                     /codon_start=1
                     /gene="HIS3"
                     /product="HIS3"
                     /label=HIS3
                     /protein_id="ABK79084.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            5431..6093
                     /gene="HIS3"
                     /label=HIS3
     promoter        complement(6565..6593)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_feature    6925..7398
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(7471..8442)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(8442..9617)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    9943..10193
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(10196..10951)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      11042..11261
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     promoter        12188..12290
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             12291..12950
                     /codon_start=1
                     /gene="CAT"
                     /product="chloramphenicol acetyl transferase"
                     /label=CAT
                     /protein_id="ABK79085.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            12291..12950
                     /gene="CAT"
                     /label=CAT
     protein_bind    13199..13232
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     promoter        13380..13398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"