LOCUS       P4_cosmid              11545 bp ds-DNA     circular     02-JAN-2023
DEFINITION  .
KEYWORDS    "marker:DR, DR, CmR"
FEATURES             Location/Qualifiers
     terminator      1..61
                     /label="termintor L3S2P21"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    62..101
                     /label="BBa_J72152"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: BBa_J72152"
     misc_feature    62..101
                     /label="Start of BBa_J72152"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: Start of BBa_J72152"
     misc_feature    104..120
                     /label="BBa_J72152"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: BBa_J72152"
     misc_feature    104..120
                     /label="Start of BBa_J72152"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: Start of BBa_J72152"
     misc_feature    104..146
                     /label="CRISPR Cas9 "
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    complement(147..317)
                     /label="tracrRNA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: tracrRNA"
     misc_RNA        complement(154..232)
                     /label="tracrRNA"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="note: trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system"
     CDS             528..4634
                     /label="cas9"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 gene: cas9 transl_table: 11 translation: MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD*"
     misc_feature    4655..4786
                     /label="crRNA leader"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system"
     repeat_region   4787..4822
                     /label="DR"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: direct repeat for the Streptococcus pyogenes CRISPR/Cas system"
     misc_feature    complement(4787..4822)
                     /label="Repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: Repeat"
     misc_feature    4823..4852
                     /label="BsaI-spc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="gene: BsaI-spc"
     misc_feature    4853..4888
                     /label="Repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: Repeat"
     repeat_region   4853..4888
                     /label="DR"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: direct repeat for the Streptococcus pyogenes CRISPR/Cas system"
     terminator      4929..5023
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: note: transcription terminator from phage lambda"
     misc_feature    5024..7655
                     /label="pbbr1 cas9 fragment"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     rep_origin      5130..5901
                     /label="pBBR1 oriV"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="note: replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication"
     CDS             5902..6564
                     /label="rep"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 gene: rep product: replication protein protein_id: AFV59786.1 transl_table: 11 translation: MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLHHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR*"
     gene            5902..6564
                     /label="rep"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="gene: rep"
     misc_feature    6573..6818
                     /label="oriT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: oriT"
     oriT            6575..6683
                     /label="oriT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: incP origin of transfer"
     misc_feature    6819..6830
                     /label="part of oriT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             complement(6854..7513)
                     /label="CmR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="codon_start: 1 gene: cat note: confers resistance to chloramphenicol product: chloramphenicol acetyltransferase transl_table: 1 translation: MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA*"
     promoter        7514..7625
                     /label="cat promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    7631..7655
                     /label="attB2"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
                     /note="bound_moiety: BP Clonase™ gene: mutant version of attB note: recombination site for the Gateway® BP reaction"
     repeat_region   7656..7775
                     /label="crr 120bp direct repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: crr 120bp direct repeat"
     misc_feature    7656..7955
                     /label="crr (cis required region for replication)"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: crr (cis required region for replication)"
     source          7656..7955
                     /label="source"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="db_xref: taxon: 10680 host: Escherichia coli mol_type: genomic DNA: organism: Enterobacteria phage P4"
     repeat_region   7661..7668
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7675..7682
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7693..7700
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7736..7743
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7745..7752
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     gene            7815..7955
                     /label="P4p05"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="db_xref: GeneID: 1261089 locus_tag: P4p05"
     repeat_region   7836..7955
                     /label="crr 120bp direct repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: crr 120bp direct repeat"
     repeat_region   7841..7848
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7855..7862
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p05 note: ori type 1 repeat"
     repeat_region   7873..7880
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p05 note: ori type 1 repeat"
     repeat_region   7916..7923
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   7925..7932
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     misc_feature    7956..8730
                     /label="cosmid fragment"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     8230..8246
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#a020f0"
                     /ApEinfo_fwdcolor="#a020f0"
                     /note="note: common sequencing primer, one of multiple similar variants"
     misc_feature    8360..8664
                     /label="P4 packaging signal"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="note: P4 packaging region. Functional cos region of bacteriophage P4, surrounded by short sequences from the original P4 chromosome, as done in the work of Trigeros et al., ensuring that every required signal sequence for the packaging was present. See: 1- Ziermann, R., & Calendar, R. (1990). Characterization of the cos sites of bacteriophages P2 and P4. Gene, 96(1), 9–15. 2- Trigueros, S., Arsuaga, J., Vazquez, M. E., Sumners, D. W. & Roca, J. Novel display of knotted DNA molecules by two-dimensional gel electrophoresis. Nucleic Acids Res. 29, e67 (2001)."
     misc_feature    complement(8667..8730)
                     /label="tsid terminator"
                     /ApEinfo_revcolor="#000080"
                     /ApEinfo_fwdcolor="#000080"
                     /note="note: Rho-independent transcription termination site.Dehò, Gianni, and D. A. N. I. E. L. A. Ghisotti. ""The satellite phage P4."" The Bacteriophages (2006): 391-408."
     gene            complement(8731..9303)
                     /label="P4p14"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="db_xref: GeneID: 1261094 locus_tag: P4p14"
     CDS             complement(8731..9303)
                     /label="P4p14"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 db_xref: UniProtKB/Swiss-Prot: P05460 db_xref: GeneID: 1261094 locus_tag: P4p14 note: psu gene product (AA 1-190) product: polarity suppression family protein protein_id: NP_042044.1 transl_table: 11 translation: MESTALQQAFDTCQNNKAAWLQRKNELAAAEQEYLRLLSGEGRNVSRLDELRNIIEVRKWQVNQAAGRYIRSHEAVQHISIRDRLNDFMQQHGTALAAALAPELMGYSELTAIARNCAIQRATDALREALLSWLAKGEKINYSAQDSDILTTIGFRPDVASVDDSREKFTPAQNMIFSRKSAQLASRQSV*"
     source          8731..10608
                     /label="source"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="db_xref: taxon: 10680 host: Escherichia coli mol_type: genomic DNA: organism: Enterobacteria phage P4"
     precursor_RNA   complement(8731..10608)
                     /label="late delta regulated transcript of sid through psu"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="db_xref: GeneID: 1261100 locus_tag: P4s05 note: late delta regulated transcript of sid through psu"
     variation       9171..9171
                     /label="in mutant psu2"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p14 note: in mutant psu2 replace: t"
     variation       9171..9171
                     /label="in mutant psu3"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p14 note: in mutant psu3 replace: t"
     variation       9245..9245
                     /label="in mutant psu-am1"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p14 note: in mutant psu-am1 replace: a"
     CDS             complement(9377..9877)
                     /label="P4p13"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 db_xref: GOA: P12551 db_xref: UniProtKB/Swiss-Prot: P12551 db_xref: GeneID: 1261088 locus_tag: P4p13 note: delta gene product (AA 1-166) product: ogr/Delta-like zinc finger family protein protein_id: NP_042043.1 transl_table: 11 translation: MIYCPSC**VAHTRRAHFMDDGTKIMIAQCRNIYCSATFEASESFFSDSKDSGMEYISGKQRYRDSLTSASCGMKRPKRMLVTGYCCRRCKGLALSRTSRRLSQEVTERFYVCTDPGCGLVFKTLQTINRFIVRPVTPDELAERLHEKQELPPVRLKTQSYSLRLE*"
     gene            complement(9377..9877)
                     /label="P4p13"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="db_xref: GeneID: 1261088 locus_tag: P4p13"
     variation       9499..9499
                     /label="in mutants org1, org2, org3 and org4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p13 note: in mutants org1, org2, org3 and org4 replace: g"
     variation       9577..9577
                     /label="in mutant delta4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p13 note: in mutant delta4 replace: t"
     variation       9580..9580
                     /label="in mutant delta6"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p13 note: in mutant delta6 replace: t"
     variation       9598..9598
                     /label="in mutant delta1 and delta2"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p13 note: in mutant delta1 and delta2 replace: a"
     gene            complement(9874..10608)
                     /label="P4p12"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="db_xref: GeneID: 1261087 locus_tag: P4p12"
     CDS             complement(9874..10608)
                     /label="P4p12"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 db_xref: UniProtKB/Swiss-Prot: P05461 db_xref: GeneID: 1261087 locus_tag: P4p12 note: sid gene product (AA 1-244) product: head size determination protein sid protein_id: NP_042042.1 transl_table: 11 translation: MSDHTIPEYLQPALAQLEKARAAHLENARLMDETVTAIERAEQEKNALAQADGNDADDWRTAFRAAGGVLSDELKQRHIERVARRELVQEYDNLAVVLNFERERLKGACDSTATAYRKAHHHLLSLYAEHELEHALNETCEALVRAMHLSILVQENPLANTTGHQGYVAPEKAVMQQVKSSLEQKIKQMQISLTGEPVLRLTGLSAATLPHMDYEVAGTPAQRKVWQDKIDQQGAELKARGLLS*"
     variation       9903..9903
                     /label="in mutant sid-am1"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="locus_tag: P4p12 note: in mutant sid-am1 replace: t"
     rep_origin      10609..11105
                     /label="origin of DNA replication"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="note: origin of DNA replication"
     source          10609..11105
                     /label="source"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="db_xref: taxon: 10680 host: Escherichia coli mol_type: genomic DNA: organism: Enterobacteria phage P4"
     repeat_region   10778..10785
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10788..10795
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10831..10838
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10853..10860
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10864..10871
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10874..10881
                     /label="ori type 1 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 1 repeat"
     repeat_region   10986..10995
                     /label="ori type 2 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 2 repeat"
     repeat_region   10996..11005
                     /label="ori type 2 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 2 repeat"
     repeat_region   11006..11015
                     /label="ori type 2 repeat"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: ori type 2 repeat"
     variation       11092..11092
                     /label="in mutant vir1"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: in mutant vir1 replace: t"
     misc_feature    11106..11140
                     /label="cosmid fragment"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             11120..11137
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="codon_start: 1 product: 6xHis affinity tag transl_table: 1 translation: HHHHHH"
     misc_feature    11141..11545
                     /label="cosmid fragment"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     terminator      11141..11188
                     /label="T7 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: transcription terminator for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11223..11239)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#a020f0"
                     /ApEinfo_fwdcolor="#a020f0"
                     /note="note: common sequencing primer, one of multiple similar variants"
     protein_bind    11247..11263
                     /label="lac operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
                     /note="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-β-D-thiogalactopyranoside (IPTG)."
     promoter        complement(11271..11277)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for the E. coli lac operon"
     promoter        complement(11278..11295)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for the E. coli lac operon"
     promoter        complement(11296..11301)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for the E. coli lac operon"
     protein_bind    11316..11337
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
                     /note="bound_moiety: E. coli catabolite activator protein note: CAP binding activates transcription in the presence of cAMP."
ORIGIN
        1 CTCGGTACCA AATTCCAGAA AAGAGGCCTC CCGAAAGGGG GGCCTTTTTT CGTTTTGGTC
       61 Cgtcaagttg tcataattgg taacgaatca gacaattgaC GgcTCGATGG AGTAGCATAG
      121 AGAagtcaga gtagaataga agtatcaaaa aaagcaccga ctcggtgcca ctttttcaag
      181 ttgataacgg actagcctta ttttaacttg ctatgctgtt ttgaatggtt ccaacaagat
      241 tattttataa cttttataac aaataatcaa ggagaaattc aaagaaattt atcagccata
      301 aaacaatact taatactata gaatgataac aaaataaact actttttaaa agaattttgt
      361 gttataatct atttattatt aagtattggg taatattttt tgaagagata ttttgaaaaa
      421 gaaaaattaa agcatattaa actaatttcg gaggtcatta aaactattat tgaaatcatc
      481 aaactcatta tggatttaat ttaaactttt tattttagga ggcaaaaatg gataagaaat
      541 actcaatagg cttagatatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat
      601 ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa
      661 aaaatcttat aggggctctt ttatttgaca gtggagagac agcggaagcg actcgtctca
      721 aacggacagc tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga
      781 ttttttcaaa tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt
      841 ttttggtgga agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg
      901 aagttgctta tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt
      961 ctactgataa agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc
     1021 gtggtcattt tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat
     1081 ttatccagtt ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg
     1141 gagtagatgc taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc
     1201 tcattgctca gctccccggt gagaagaaaa atggcttatt tgggaatctc attgctttgt
     1261 cattgggttt gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac
     1321 agctttcaaa agatacttac gatgatgatt tagataattt attggcgcaa attggagatc
     1381 aatatgctga tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata
     1441 tcctaagagt aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct
     1501 acgatgaaca tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag
     1561 aaaagtataa agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg
     1621 ggggagctag ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg
     1681 gtactgagga attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct
     1741 ttgacaacgg ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa
     1801 gacaagaaga cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga
     1861 cttttcgaat tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga
     1921 tgactcggaa gtctgaagaa acaattaccc catggaattt tgaagaagtt gtcgataaag
     1981 gtgcttcagc tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg
     2041 aaaaagtact accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga
     2101 caaaggtcaa atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga
     2161 agaaagccat tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa
     2221 aagaagatta tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag
     2281 atagatttaa tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag
     2341 attttttgga taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct
     2401 tatttgaaga tagggagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg
     2461 ataaggtgat gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa
     2521 aattgattaa tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat
     2581 cagatggttt tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta
     2641 aagaagacat tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg
     2701 caaatttagc tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg
     2761 atgaattggt caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac
     2821 gtgaaaatca gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg
     2881 aagaaggtat caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc
     2941 aattgcaaaa tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg
     3001 accaagaatt agatattaat cgtttaagtg attatgatgt cgatcacatt gttccacaaa
     3061 gtttccttaa agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg
     3121 gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac
     3181 aacttctaaa cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac
     3241 gtggaggttt gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc
     3301 gccaaatcac taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg
     3361 aaaatgataa acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg
     3421 acttccgaaa agatttccaa ttctataaag tacgtgagat taacaattac catcatgccc
     3481 atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg
     3541 aatcggagtt tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt
     3601 ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact
     3661 tcttcaaaac agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa
     3721 ctaatgggga aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca
     3781 aagtattgtc catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat
     3841 tctccaagga gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag
     3901 actgggatcc aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag
     3961 tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag
     4021 ggatcacaat tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta
     4081 aaggatataa ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg
     4141 agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg
     4201 agctggctct gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt
     4261 tgaagggtag tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt
     4321 atttagatga gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg
     4381 ccaatttaga taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac
     4441 aagcagaaaa tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta
     4501 aatattttga tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa gttttagatg
     4561 ccactcttat ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc
     4621 taggaggtga ctgaagtata ttttagatga agattatttc ttaataacta aaaatatggt
     4681 ataatactct taataaatgc agtaatacag gggcttttca agactgaagt ctagctgaga
     4741 caaatagtgc gattacgaaa ttttttagac aaaaatagtc tacgaggttt tagagctatg
     4801 ctgttttgaa tggtcccaaa actgagacca gtctcggaag ctcaaaggtc tcgttttaga
     4861 gctatgctgt tttgaatggt cccaaaactt cagcacactg agacttgttg agtGGATCCG
     4921 AGGCTTGGAT TCTCACCaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     4981 agatggagtt ctgaggtcat tactggatct atcaacagga gtcctagaca gctgggcgcg
     5041 ccccccctac gggcttgctc tccgggcttc gccctgcgcg gtcgctgcgc tcccttgcca
     5101 gcccgtggat atgtggacga tggccgcgag cggccaccgg ctggctcgct tcgctcggcc
     5161 cgtggacaac cctgctggac aagctgatgg acaggctgcg cctgcccacg agcttgacca
     5221 cagggattgc ccaccggcta cccagccttc gaccacatac ccaccggctc caactgcgcg
     5281 gcctgcggcc ttgccccatc aattttttta attttctctg gggaaaagcc tccggcctgc
     5341 ggcctgcgcg cttcgcttgc cggttggaca ccaagtggaa ggcgggtcaa ggctcgcgca
     5401 gcgaccgcgc agcggcttgg ccttgacgcg cctggaacga cccaagccta tgcgagtggg
     5461 ggcagtcgaa gggcgaagcc cgcccgcctg ccccccgagc ctcacggcgg cgagtgcggg
     5521 ggttccaagg gggcagcgcc accttgggca aggccgaagg ccgcgcagtc gatcaacaag
     5581 ccccggaggg gccacttttt gccggagggg gagccgcgcc gaaggcgtgg gggaaccccg
     5641 caggggtgcc cttctttggg caccaaagaa ctagatatag ggcgaaatgc gaaagactta
     5701 aaaatcaaca acttaaaaaa ggggggtacg caacagctca ttgcggcacc ccccgcaata
     5761 gctcattgcg taggttaaag aaaatctgta attgactgcc acttttacgc aacgcataat
     5821 tgttgtcgcg ctgccgaaaa gttgcagctg attgcgcatg gtgccgcaac cgtgcggcac
     5881 ccctaccgca tggagataag catggccacg cagtccagag aaatcggcat tcaagccaag
     5941 aacaagcccg gtcactgggt gcaaacggaa cgcaaagcgc atgaggcgtg ggccgggctt
     6001 attgcgagga aacccacggc ggcaatgctg ctgcatcacc tcgtggcgca gatgggccac
     6061 cagaacgccg tggtggtcag ccagaagaca ctttccaagc tcatcggacg ttctttgcgg
     6121 acggtccaat acgcagtcaa ggacttggtg gccgagcgct ggatctccgt cgtgaagctc
     6181 aacggccccg gcaccgtgtc ggcctacgtg gtcaatgacc gcgtggcgtg gggccagccc
     6241 cgcgaccagt tgcgcctgtc ggtgttcagt gccgccgtgg tggttgatca cgacgaccag
     6301 gacgaatcgc tgttggggca tggcgacctg cgccgcatcc cgaccctgta tccgggcgag
     6361 cagcaactac cgaccggccc cggcgaggag ccgcccagcc agcccggcat tccgggcatg
     6421 gaaccagacc tgccagcctt gaccgaaacg gaggaatggg aacggcgcgg gcagcagcgc
     6481 ctgccgatgc ccgatgagcc gtgttttctg gacgatggcg agccgttgga gccgccgaca
     6541 cgggtaacgc tgccgcgccg gtagggccgg cctacggcca gcctcgcaga gcaggattcc
     6601 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     6661 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     6721 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     6781 ctataatgac cccgaagcag ggttatgcag cggaaaagga caaaagtcaa Atgcaagcga
     6841 gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     6901 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     6961 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     7021 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     7081 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     7141 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     7201 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     7261 atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc
     7321 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     7381 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     7441 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     7501 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     7561 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaaAG
     7621 ATCCgGATCA ACCACTTTGT ACAAGAAAGC TGGGTccgaa ggtgaacaat ccactgttca
     7681 cccttcaccg tatattcacc cgttatcaca ctgaaattaa aagagaaaaa cgaaaggtga
     7741 acagtgtgaa caatcaaatc aaaaaaaact tttttcttcc tgagtgattt cagtgcggag
     7801 gattaatcac cggtatgagt cacatcggca gaatgccgga ggtgaagaat cgaatgttca
     7861 cccttcaccc attattcacc acctatcact ctgaaataaa aggagaaaac agaaaggtga
     7921 acagtgtgaa cagttctttc gaaaaaaaat tttttGGCGC GTCAGCGGGT GTTGGCGGGT
     7981 GTCGGGGCTG GCTTAACTAT GCGGCATCAG AGCAGATTGT ACTGAGAGTG CACCATATGC
     8041 GGTGTGAAAT ACCGCACAGA TGCGTAAGGA GAAAATACCG CATCAGGCGC CATTCGCCAT
     8101 TCAGGCTGCG CAACTGTTGG GAAGGGCGAT CGGTGCGGGC CTCTTCGCTA TTACGCCAGC
     8161 TGGCGAAAGG GGGATGTGCT GCAAGGCGAT TAAGTTGGGT AACGCCAGGG TTTTCCCAGT
     8221 CACGACGTTG TAAAACGACG GCCAGTGAAA TTATTAACCT AGGCTGACTT CATCCGCACA
     8281 CACGAGCCGC GCCCCTTGCG CGACGGGCTG GCGGTATTGG CGGCGCTGAA AGAAACCGCA
     8341 AGAATTCTTA AGGATCTTGC GACACTGACA AGAACCACCT TACATCAATC TATTTTTTGC
     8401 ATATTTTTCA GGATAGTGCC ATCAAAAAAG ACAGTTTTGG ACCATAGGCC ATAGATTTTC
     8461 TTCATTATGC TCCAATGTAT TAACGAGGTA TGTACTGAAG GCACTTTGAC TACACAAATA
     8521 ATTAACTAAC TTGTCGGTTT GCGCGCGGTG CTTTCCCCGC CTCGCCCGCC CGCTTTGCGG
     8581 GGCGGTTTTA ATGCAGTTGC ACTGACACGC TCAGACCGCG CCGGGAATGG CGCGGTTTGC
     8641 AGAAAATGAG GCAGGAAAAC GCATGCAAAG CCATGCACCT TATCGATGCA TGGCTTTTTT
     8701 CAGTAAAAAC GGGCGGATTT TCGGGGAATT ttacactgac tgacgtgatg ccagttgcgc
     8761 acttttacgc gaaaaaatca tgttctgcgc aggggtgaat ttttcacggc tgtcatccac
     8821 cgaagccacg tcaggcctga atccgatggt cgttaaaatg tcgctatcct gtgcggaata
     8881 attaattttt tcacccttcg caagccagga cagaagggct tcacgcaggg catctgtggc
     8941 acgctgtatg gcacagtttc gggcaatggc cgtcagctca ctgtagccca tcagctccgg
     9001 tgccagtgcg gccgccagtg ctgtgccgtg ctgctgcata aaatcattca gccggtcgcg
     9061 gatgctgatg tgctgaacgg cttcatgcga acgaatataa cgaccggcgg cctgattcac
     9121 ctgccatttt ctgacttcga taatattgcg taattcgtcc aggcgactga cgtttctgcc
     9181 ttctcctgac agaagccgca gatattcctg ttcggccgct gccagctcat ttttgcgttg
     9241 cagccatgct gctttgttat tctgacaggt gtcaaaggcc tgctgtaagg ctgtgctttc
     9301 catcgttatc tctttctcat catgctgaag aataaaaata cggtgtgcgg cgacggccgg
     9361 tgttaaccgg cagccctcat tccagacgca gcgaatatga ttgtgttttt aaccgtactg
     9421 gcggcagttc ctgtttttca tgcaggcgtt ctgccagttc gtccggcgtg accgggcgga
     9481 caatgaagcg gttgatggtc tgaagcgttt taaacaccag accacagccc ggatccgtgc
     9541 acacataaaa acgctcggtg acttcctgag acagacgccg cgatgttctt gacagtgcaa
     9601 ggcctttaca tctccgacaa caatatccgg taacaagcat tcttttcggg cgtttcatac
     9661 cgcaggaggc tgacgtcagt gaatcgcggt atctctgttt gcctgaaatg tattccattc
     9721 ctgaatcttt actgtcagag aaaaagcttt cactcgcttc aaatgtcgca gagcaataaa
     9781 tattccggca ctgtgcaatc attatcttgg tgccatcgtc catgaaatgt gcgcgacggg
     9841 tgtgagcaac atgtccacac gacggacagt aaatcatgac agcagccctc tggccttaag
     9901 ctctgctccc tgctggtcta ttttgtcctg ccacaccttg cgctgtgccg gtgtgcctgc
     9961 cacctcataa tccatgtgcg ggagtgttgc cgctgacagt ccggtcagcc ggagaaccgg
    10021 ctcgccggtg aggctgattt gcatctgttt aattttctgt tccagcgatg atttcacctg
    10081 ctgcatgaca gccttttccg gtgcgacgta gccctgatgg ccggtggtgt tggcgagcgg
    10141 attttcctgt accagaatgc tcagatgcat tgcccggaca agcgcctcac aggtttcatt
    10201 cagggcgtgt tccagctcat gctctgcata cagactcaga aggtgatgat gtgccttccg
    10261 gtaggcggtg gccgtgctgt cacacgcccc tttcaggcgt tcacgttcga aattcagcac
    10321 cacggccaga ttgtcatatt cctgtaccag ctcccggcgt gccacgcgct caatgtggcg
    10381 ctgtttcagc tcgtcgctca ggacaccacc ggctgcacga aaggccgtgc gccagtcgtc
    10441 agcgtcgttt ccgtcggcct gcgccagcgc atttttttcc tgctctgccc gttcaatggc
    10501 cgtgacagtc tcatccatca ggcgggcgtt ctcaagatgg gcggctctgg ccttttccag
    10561 ttgtgccagt gcgggttgca gatattcagg gatagtgtgg tcagacattt tccggctcct
    10621 cgtcacttca ggttaagaaa attgtgacgt acaccggaca acaacacgac gcattgcaga
    10681 tgtgccagcc ctgacacagg agactcatcc tcagaccggc aagccaggaa aaggtcgcag
    10741 gaaaaaccgg cttactgttt gtttttttat attttactgt tcacctctgt tcaccttaat
    10801 aaaaaagata agtaatacag taagttaaag ggtgaacaat cgcagtaatg actgttcacc
    10861 gtctgttcac cactgttcac ccgttgatgg gctttttgtg ctgtttacta ctgtttgttt
    10921 ttattaattc gccgggaatg aataagaaga aacgatttgt atttcactat aaaaattaca
    10981 aattgcttta agtggcttta aatgacttta agtggagatg aacaaaaaac acacagccat
    11041 tgtaagacag cctgaacaaa tcccccctgt tgcgtctgct gaaaatattc acaaaataaa
    11101 gcgctCAGCA CAGGCAGCAC ATCATCATCA CCACCACTAA CTAGCATAAC CCCTTGGGGC
    11161 CTCTAAACGG GTCTTGAGGG GTTTTTTGAT CCCAATGGCG CGCCGAGCTT GGCTCGAGCA
    11221 TGGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CAACATACGA
    11281 GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT
    11341 GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAATGA
    11401 ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC
    11461 ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG
    11521 GTAATACGGT TATCCACAGA ATCAG
//