LOCUS       Exported                4759 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pT181mcs, complete sequence.
ACCESSION   JX893516
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4759)
  AUTHORS   Coelho MLV.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2012) Instituto de Microbiologia, Universidade 
            Federal do Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho, 373. 
            Edificio do Centro de Ciencias da Saude, Bloco I, Cidade 
            Universitaria, Rio de Janeiro, RJ 21941-902, Brazil
REFERENCE   2  (bases 1 to 4759)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4759
                     /organism="Cloning vector pT181mcs"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: m13f, fwd_seq:
                     gttttcccagtcacgacgttgta, rev_name: m13r, rev_seq: 
                     caggaaacagctatgacc"
                     /db_xref="taxon:1238575"
     CDS             1046..1843
                     /codon_start=1
                     /gene="tetL"
                     /product="tetracycline resistance protein"
                     /label=tetL
                     /protein_id="AFU76897.1"
                     /translation="MVLNVSLPDIANHFNTTPGITNWVNTAYMLTFSIGTAVYGKLSDY
                     INIKKLLIIGISLSCLGSLIAFIGHNHFFILIFGRLVQGVGSAAFPSLIMVVVARNITR
                     KKQGKAFGFIGSIVALGEGLGPSIGGIIAHYIHWSYLLILPMITIVTIPFLIKVMVPGK
                     STKNTLDIVGIVLMSISIICFMLFTTNYNWTFLILFTIFFVIFIKHISRVSNPFINPKL
                     GKNIPFMLGLFSGWANIFYSSWFYINGALYDENYLSCKCSDNR"
     gene            1046..1843
                     /gene="tetL"
                     /label=tetL
     CDS             1788..2258
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AFU76898.1"
                     /translation="MVPYMMKTIYHVNVATIGNSVIFPGTMSVIVFGYFGGFLVDRKGS
                     LFVFILGSLSISISFLTIAFFVEFSMWLTTFMFIFVMGELSFTKTVISKIVSSSLSEEE
                     VASGMSLLNFTSFLSEGTGIAIVGGLLSLQLINRKLVLEFINYSSGVYSNIL"
     gene            1788..2258
                     /gene="tet"
                     /label=tet
     CDS             2521..2952
                     /codon_start=1
                     /gene="mob_pre_N"
                     /product="truncated mobilization protein"
                     /label=mob_pre_N
                     /protein_id="AFU76901.1"
                     /translation="MSYSIVRVSKVKSGTNTTGIQKHVQRENNNYENEDIDHSKTYLNY
                     DLVNANKQNFNNLIDEKIEQNYTGKRKIRTDAIKHIDGLITSDNDFFDNQTPEDTKQFF
                     EYAKEFLEQEYGKDNLLYATVHMDEKTPHWRKGDVLQGD"
     gene            2521..2952
                     /gene="mob_pre_N"
                     /label=mob_pre_N
                     /note="disrupted by insertion of pUC19 MCS"
     CDS             complement(2840..3079)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="AFU76899.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASVVFFRPCELLHIINYLYRILVLKTL"
     gene            complement(2840..3079)
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     2989..3005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3006..3062
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3075..3091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3099..3115
                     /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(3123..3153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             3168..4082
                     /codon_start=1
                     /gene="mob_pre_C"
                     /product="truncated mobilization protein"
                     /label=mob_pre_C
                     /protein_id="AFU76900.1"
                     /translation="MSELTHINCVALTARFPVGKPVVPVHYGVVPITDDGRLSAKEVVG
                     NKKALTAFQDRFNEHVKQRGYGLERGQSRQVTNAKHEQISQYKQKTEYHKQEYERESQK
                     TDHIKQKNDKLMQEYQKSLNTLKKPINVPYEQETEKVGGLFSKEIQEAGNVVISQKDFN
                     EFQKQIKAAQDISEDYEYIKSGRALDDKDKEIREKDDLLNKAVERIENADDNFNQLYEN
                     AKPLKENIEIALKLLKILLKELERVLGRNTFAERVNKLTEDEPKLNGLAGNLDKKMNPE
                     LYSEQEQQQEQQKNQKRDRGMHL"
     gene            3168..4082
                     /gene="mob_pre_C"
                     /label=mob_pre_C
                     /note="disrupted by insertion of pUC19 MCS; translated only
                     if an active promoter is cloned into MCS"
     protein_bind    3168..3189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             4609..794
                     /codon_start=1
                     /gene="rep_trans"
                     /product="Rep"
                     /label=rep_trans
                     /note="plasmid replication initiation protein"
                     /protein_id="AFU76902.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEEDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     gene            4609..794
                     /gene="rep_trans"
                     /label=rep_trans