LOCUS       Exported               11365 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC82, complete sequence.
ACCESSION   FJ389187
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11365)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Growth rate modulates the selective advantage of recurrent 
            transposition mutations arising from experimental populations grown 
            in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11365)
  AUTHORS   Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 11365)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11365
                     /organism="Cloning vector pHC82"
                     /mol_type="other DNA"
                     /db_xref="taxon:646109"
     CDS             complement(1..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin-resistance protein"
                     /protein_id="ACR45012.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1..861)
                     /gene="bla"
                     /label=bla
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1053..4376
                     /note="icuAB.CM1059 allele of Methylobacterium extorquens 
                     AM1"
     CDS             2053..3267
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /note="transposase of ISMex4"
                     /protein_id="ACR45013.1"
                     /translation="MQIGEKLAAVIPDRRDPSRIVHPLPEILLARILAIACGYEDADDL
                     DHLRVDPAFKLACGRLPDSGRDLMSQPTVSRLENTPGLRDLIRLGRVLVDLYCASYAAP
                     PAAVTLDIDDTCDVVHGQQQLSLFNAHHDERCFLPIHVYDTATSRPVAMILRPGKTPSG
                     REVRGHLRRLVRAIRRHWPTTAITIRGDGHYGRPEAMDWCEDNDVAYVFGLTGTKSLTA
                     KVEPTADHIRVERALSNTDAVRGFAETTHAAKSWRQHRRVAARIEATRLGLDIRYVVTN
                     IATGTPEWLYAELYCARGQAENLIKLHKGQLASDRTSCRHPAANQMRLVLHTAAYWLML
                     TVRDAIPAPHRLAKAEFATIRLRLLKLGARIHETTARVRLAFAAACPEADLLRHIAGAL
                     APAPA"
     oriT            4887..5625
                     /label=origin of transfer
                     /note="origin of transfer"
     CDS             complement(4977..5348)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5381..5490
                     /note="incP origin of transfer"
     promoter        6127..6155
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             6203..7393
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline-resistance protein"
                     /label=tet
                     /protein_id="ACR45014.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            6203..7393
                     /gene="tet"
                     /label=tet
     CDS             complement(7571..8230)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(8328..9749)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="ACR45015.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(8328..9749)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(9750..10195)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(10565..11120)
                     /direction=LEFT
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(10607..11195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"