LOCUS       SEGA072                4746 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12908.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4746)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PT7-medium-YebF-sfGFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4746
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          1868..1930
                     /organism="Escherichia coli str. K-12 substr. MG1655"
                     /strain="K-12"
                     /sub_strain="MG1655"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:511145"
     CDS             1..528
                     /codon_start=3
                     /label=glmS
                     /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI
                     ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR
                     ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD
                     QPRNLAKSVTVE"
     misc_feature    538..594
                     /label=L3S2P11 Terminator
     CDS             complement(604..1686)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1687..1764)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1795..1813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1814..1838
                     /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)."
     RBS             1850..1861
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1862..1876
                     /label=TIR
     gene            1868..1930
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /label=yebF
                     /db_xref="EcoGene:EG11807"
                     /db_xref="GeneID:946363"
     CDS             1868..1930
                     /codon_start=1
                     /transl_table=11
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /product="extracellular Colicin M immunity family protein"
                     /label=yebF
                     /db_xref="GI:145698278"
                     /db_xref="ASAP:ABE-0006141"
                     /db_xref="UniProtKB/Swiss-Prot:P33219"
                     /db_xref="EcoGene:EG11807"
                     /db_xref="GeneID:946363"
                     /protein_id="NP_416361.2"
                     /translation="MKKRGAFLGLLLVSACASVFA"
     CDS             1931..2221
                     /codon_start=1
                     /label=YebF
                     /translation="ANNETSKSVTFPKCEDLDAAGIAASVKRDYQQNRVARWADDQKIV
                     GQADPVAWVSLQDIQGKDDKWSVPLTVRGKSADIHYQVSVDCKAGMAEYQRR"
     CDS             2222..2932
                     /codon_start=1
                     /label=sfGFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2936..2984
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2995..3023
                     /label=Promoter 3BCD2
     CDS             3063..4253
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    4234..4283
                     /label=HR3
     terminator      4264..4304
                     /label=rrnC
     misc_feature    4305..4354
                     /label=homology
     CDS             4499..4746
                     /codon_start=1
                     /label=pstS
                     /translation="MKVMRTTVATVVAATLSMSAFSVFAEASLTGAGATFPAPVYAKWA
                     DTYQKETGNKVNYQGIGSSGGVKQIIANTVDFGASDA"
ORIGIN
        1 gtgcatggtc tgcaggcgct gccgagccgt attgagcaga tgctgtctca ggacaaacgc
       61 attgaagcgc tggcagaaga tttctctgac aaacatcacg cgctgttcct gggccgtggc
      121 gatcagtacc caatcgcgct ggaaggcgca ttgaagttga aagagatctc ttacattcac
      181 gctgaagcct acgctgctgg cgaactgaaa cacggtccgc tggcgctaat tgatgccgat
      241 atgccggtta ttgttgttgc accgaacaac gaattgctgg aaaaactgaa atccaacatt
      301 gaagaagttc gcgcgcgtgg cggtcagttg tatgtcttcg ccgatcagga tgcgggtttt
      361 gtaagtagcg ataacatgca catcatcgag atgccgcatg tggaagaggt gattgcaccg
      421 atcttctaca ccgttccgct gcagctgctg gcttaccatg tcgcgctgat caaaggcacc
      481 gacgttgacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatctc
      541 ggtaccaaat tccagaaaag agacgctttc gagcgtcttt tttcgttttg gtccgcgttg
      601 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      661 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
      721 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
      781 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
      841 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
      901 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
      961 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     1021 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     1081 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     1141 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     1201 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     1261 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     1321 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     1381 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     1441 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     1501 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     1561 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     1621 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     1681 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     1741 ggttttgcgc cattcgatgg tgtcggatcg agatcgatct cgatcccgcg aaattaatac
     1801 gactcactat aggggaattg tgagcggata acaattccta ctagagatta aagaggagaa
     1861 agactggatg aaaaagagag gggcgttttt agggctgttg ttggtttctg cctgcgcatc
     1921 agttttcgct gcgaataacg aaacgagcaa gagcgtcacc ttcccgaagt gcgaggatct
     1981 ggacgcagcg ggcattgctg ccagcgtgaa acgtgactac caacagaacc gcgtggctcg
     2041 ttgggcagac gaccagaaga tcgtcggtca agcggacccg gttgcctggg tttccctgca
     2101 agatatccag ggcaaagatg ataaatggag cgtcccgctg accgttcgtg gtaagagcgc
     2161 ggacattcac tatcaggtga gcgtcgattg caaagccggc atggcggagt accaacgccg
     2221 ccgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
     2281 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
     2341 actgacgctg aagttcatct gtactactgg taaactgccg gtaccttggc cgactctggt
     2401 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccaca tgaagcagca
     2461 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
     2521 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
     2581 ccgcattgag ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct
     2641 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
     2701 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
     2761 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
     2821 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcaca tggttctgct
     2881 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccaat
     2941 tattgaaggc cgctaacgcg gccttttttt gtttctggtc tcccaaaaat ttatttgctt
     3001 attaatcatc cggctcgtat aatgtgtgga gggcccaagt tcacttaaaa aggagaatag
     3061 ttatgaaatc gaacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag
     3121 gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc cattccgaca
     3181 gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa tttctatgcg
     3241 cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc
     3301 tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg tggattctct
     3361 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
     3421 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
     3481 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccctgc
     3541 acgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
     3601 taatgcagga gtcgcataag ggagagcgcc gtccgatgcc cttgagagcc ttcaacccag
     3661 tcagctcctt ccggtgggcg cggggcatga ccattgtggc cgcacttatg actgtcttct
     3721 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     3781 gctttcgctg gagcgcgacg atgagcggcc tgtcgcttgc ggtattcgga atcttgcacg
     3841 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     3901 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     3961 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     4021 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     4081 tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg gcgatttatg
     4141 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     4201 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgagacaatt
     4261 gtccagataa aaaaaatcct tagctttcgc taaggatgat ttctaaaatt tttctgtcat
     4321 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttaaa tcagactgaa
     4381 gactttatct ctctgtcata aaactgtcat attccttaca tataactgtc acctgtttgt
     4441 cctattttgc ttctcgtagc caacaaacaa tgctttatga atcctcccag gagacattat
     4501 gaaagttatg cgtaccaccg tcgcaactgt tgtcgccgcg accttatcga tgagtgcttt
     4561 ctctgtgttt gcagaagcaa gcctgacagg tgcaggtgca accttccctg cgccggtgta
     4621 tgccaaatgg gctgacactt accagaaaga aaccggtaat aaagttaact accagggtat
     4681 cggttcttcc ggtggcgtaa aacagattat cgctaatacc gttgattttg gtgcctctga
     4741 cgcgcc
//
