LOCUS       SEGA025                4711 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12861.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4711)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: Ptrc-high-Ubiquitin-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4711
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     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
     CDS             1878..2177
                     /codon_start=1
                     /label=Ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPGSAHHHHHHHHHHAGSSDTIENVKAK
                     IQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQEKSTLELVLELQNETQSATSDA"
     CDS             1944..1973
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2187..2897
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006) ?"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2901..2949
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2960..2988
                     /label=Promoter 3BCD2
     CDS             3028..4218
                     /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    4199..4248
                     /label=HR3
     terminator      4229..4269
                     /label=rrnC
     misc_feature    4270..4319
                     /label=homology
     CDS             4464..4711
                     /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 acattgtatg cagatattcg tgaaaacact gaccggcaaa accattaccc
     1921 tggaggtgga gcctggcagc gcccatcatc atcaccatca ccatcaccat catgctggta
     1981 gcagcgatac gatcgaaaac gtcaaagcca aaattcagga caaagagggc attccgcctg
     2041 atcaacaacg tctgatcttc gccggaaaac agctggagga tggccgtaca ctgagtgact
     2101 ataacattca agagaaatcg acactggaac tggttctgga gctgcagaat gaaacccagt
     2161 cagcaacttc tgacgcttct ggcggtagca aaggagaaga acttttcact ggagttgtgc
     2221 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     2281 gtgaaggtga tgctacatac ggaaaactca cccttaaatt tatttgcact actggaaaac
     2341 tacctgttcc atggccaaca cttgtcacta ctctgaccta tggcgttcaa tgcttttccc
     2401 gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     2461 tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca
     2521 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag
     2581 atggaaacat tctcggacac aaactagagt acaactataa ctcacacaat gtatacatca
     2641 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag
     2701 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     2761 tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg
     2821 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
     2881 tggatgagct ctacaaatga ccaattattg aaggccgcta acgcggcctt tttttgtttc
     2941 tggtctccca aaaatttatt tgcttattaa tcatccggct cgtataatgt gtggagggcc
     3001 caagttcact taaaaaggag aatagttatg aaatcgaaca atgcgctcat cgtcatcctc
     3061 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
     3121 ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta
     3181 tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc
     3241 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg
     3301 accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat caccggcgcc
     3361 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     3421 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     3481 ggactgttgg gcgccatctc cctgcacgca ccattccttg cggcggcggt gctcaacggc
     3541 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgccgtccg
     3601 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgaccatt
     3661 gtggccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3721 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgag cggcctgtcg
     3781 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     3841 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     3901 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     3961 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     4021 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga
     4081 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     4141 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     4201 cgggccacct cgacctgaga caattgtcca gataaaaaaa atccttagct ttcgctaagg
     4261 atgatttcta aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat
     4321 caacgcttat ttaaatcaga ctgaagactt tatctctctg tcataaaact gtcatattcc
     4381 ttacatataa ctgtcacctg tttgtcctat tttgcttctc gtagccaaca aacaatgctt
     4441 tatgaatcct cccaggagac attatgaaag ttatgcgtac caccgtcgca actgttgtcg
     4501 ccgcgacctt atcgatgagt gctttctctg tgtttgcaga agcaagcctg acaggtgcag
     4561 gtgcaacctt ccctgcgccg gtgtatgcca aatgggctga cacttaccag aaagaaaccg
     4621 gtaataaagt taactaccag ggtatcggtt cttccggtgg cgtaaaacag attatcgcta
     4681 ataccgttga ttttggtgcc tctgacgcgc c
//
