LOCUS       SEGA035                4756 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12871.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4756)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: Ptrc-low-YebF-sfGFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4756
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          1878..1940
                     /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
     misc_feature    1795..1824
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        1795..1824
                     /label=The trc promoter combines the -35 region from t
                     /label=The trc promoter combines the -35 region from the
                     /note="The trc promoter combines the -35 region from the 
                     trp promoter and the -10 region from the lacUV5 promoter"
     -35_signal      1795..1800
                     /gene="trp"
                     /label=trp
     misc_feature    1795..1800
                     /label=-35
     -10_signal      1818..1824
                     /gene="lacUV5"
                     /label=lacUV5
     misc_feature    1818..1824
                     /label=-10
     misc_feature    1825..1848
                     /label=lacO
     RBS             1860..1871
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1872..1886
                     /label=TIR
     gene            1878..1940
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /label=yebF
                     /db_xref="EcoGene:EG11807"
                     /db_xref="GeneID:946363"
     CDS             1878..1940
                     /codon_start=1
                     /transl_table=11
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /product="extracellular Colicin M immunity family protein"
                     /label=YebFss
                     /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             1941..2231
                     /codon_start=1
                     /label=YebF
                     /translation="ANNETSKSVTFPKCEDLDAAGIAASVKRDYQQNRVARWADDQKIV
                     GQADPVAWVSLQDIQGKDDKWSVPLTVRGKSADIHYQVSVDCKAGMAEYQRR"
     CDS             2232..2942
                     /codon_start=1
                     /label=sfGFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2946..2994
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3005..3033
                     /label=Promoter 3BCD2
     CDS             3073..4263
                     /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    4244..4293
                     /label=HR3
     terminator      4274..4314
                     /label=rrnC
     misc_feature    4315..4364
                     /label=homology
     CDS             4509..4756
                     /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 aaatttgaca
     1801 attaatcatc cggctcgtat aatggaattg tgagcggata acaattccta ctagagatta
     1861 aagaggagaa accctcgatg aagaaaagag gggcgttttt agggctgttg ttggtttctg
     1921 cctgcgcatc agttttcgct gcgaataacg aaacgagcaa gagcgtcacc ttcccgaagt
     1981 gcgaggatct ggacgcagcg ggcattgctg ccagcgtgaa acgtgactac caacagaacc
     2041 gcgtggctcg ttgggcagac gaccagaaga tcgtcggtca agcggacccg gttgcctggg
     2101 tttccctgca agatatccag ggcaaagatg ataaatggag cgtcccgctg accgttcgtg
     2161 gtaagagcgc ggacattcac tatcaggtga gcgtcgattg caaagccggc atggcggagt
     2221 accaacgccg ccgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac
     2281 tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa
     2341 ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg gtaccttggc
     2401 cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccaca
     2461 tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga
     2521 tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata
     2581 ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaagacggc aatatcctgg
     2641 gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa
     2701 aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc
     2761 tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca
     2821 atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcaca
     2881 tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca
     2941 aatgaccaat tattgaaggc cgctaacgcg gccttttttt gtttctggtc tcccaaaaat
     3001 ttatttgctt attaatcatc cggctcgtat aatgtgtgga gggcccaagt tcacttaaaa
     3061 aggagaatag ttatgaaatc gaacaatgcg ctcatcgtca tcctcggcac cgtcaccctg
     3121 gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc
     3181 cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa
     3241 tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg
     3301 ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg
     3361 tggattctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     3421 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     3481 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     3541 atctccctgc acgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     3601 ggctgcttcc taatgcagga gtcgcataag ggagagcgcc gtccgatgcc cttgagagcc
     3661 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ccattgtggc cgcacttatg
     3721 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     3781 ggcgaggacc gctttcgctg gagcgcgacg atgagcggcc tgtcgcttgc ggtattcgga
     3841 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     3901 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     3961 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     4021 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     4081 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg
     4141 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     4201 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     4261 tgagacaatt gtccagataa aaaaaatcct tagctttcgc taaggatgat ttctaaaatt
     4321 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttaaa
     4381 tcagactgaa gactttatct ctctgtcata aaactgtcat attccttaca tataactgtc
     4441 acctgtttgt cctattttgc ttctcgtagc caacaaacaa tgctttatga atcctcccag
     4501 gagacattat gaaagttatg cgtaccaccg tcgcaactgt tgtcgccgcg accttatcga
     4561 tgagtgcttt ctctgtgttt gcagaagcaa gcctgacagg tgcaggtgca accttccctg
     4621 cgccggtgta tgccaaatgg gctgacactt accagaaaga aaccggtaat aaagttaact
     4681 accagggtat cggttcttcc ggtggcgtaa aacagattat cgctaatacc gttgattttg
     4741 gtgcctctga cgcgcc
//
