LOCUS       SEGA059                4701 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12895.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4701)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PT7-low-Ubiquitin-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4701
                     /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
     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
     CDS             1868..2167
                     /codon_start=1
                     /label=Ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPGSAHHHHHHHHHHAGSSDTIENVKAK
                     IQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQEKSTLELVLELQNETQSATSDA"
     CDS             1934..1963
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2177..2887
                     /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    2891..2939
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2950..2978
                     /label=Promoter 3BCD2
     CDS             3018..4208
                     /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    4189..4238
                     /label=HR3
     terminator      4219..4259
                     /label=rrnC
     misc_feature    4260..4309
                     /label=homology
     CDS             4454..4701
                     /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 aggggggatg caaattttcg tgaaaacact gaccggcaaa accattaccc tggaggtgga
     1921 gcctggcagc gcccatcatc atcaccatca ccatcaccat catgctggta gcagcgatac
     1981 gatcgaaaac gtcaaagcca aaattcagga caaagagggc attccgcctg atcaacaacg
     2041 tctgatcttc gccggaaaac agctggagga tggccgtaca ctgagtgact ataacattca
     2101 agagaaatcg acactggaac tggttctgga gctgcagaat gaaacccagt cagcaacttc
     2161 tgacgcttct ggcggtagca aaggagaaga acttttcact ggagttgtgc caattcttgt
     2221 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
     2281 tgctacatac ggaaaactca cccttaaatt tatttgcact actggaaaac tacctgttcc
     2341 atggccaaca cttgtcacta ctctgaccta tggcgttcaa tgcttttccc gttatccgga
     2401 tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg
     2461 cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg
     2521 tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag atggaaacat
     2581 tctcggacac aaactagagt acaactataa ctcacacaat gtatacatca cggcagacaa
     2641 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggttccgt
     2701 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     2761 agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga
     2821 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct
     2881 ctacaaatga ccaattattg aaggccgcta acgcggcctt tttttgtttc tggtctccca
     2941 aaaatttatt tgcttattaa tcatccggct cgtataatgt gtggagggcc caagttcact
     3001 taaaaaggag aatagttatg aaatcgaaca atgcgctcat cgtcatcctc ggcaccgtca
     3061 ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata
     3121 tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga
     3181 tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag
     3241 tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg
     3301 tcctgtggat tctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     3361 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     3421 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     3481 gcgccatctc cctgcacgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     3541 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgccgtccg atgcccttga
     3601 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgaccatt gtggccgcac
     3661 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     3721 ttttcggcga ggaccgcttt cgctggagcg cgacgatgag cggcctgtcg cttgcggtat
     3781 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     3841 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     3901 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     3961 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     4021 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga ccgctgatcg
     4081 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     4141 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     4201 cgacctgaga caattgtcca gataaaaaaa atccttagct ttcgctaagg atgatttcta
     4261 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     4321 ttaaatcaga ctgaagactt tatctctctg tcataaaact gtcatattcc ttacatataa
     4381 ctgtcacctg tttgtcctat tttgcttctc gtagccaaca aacaatgctt tatgaatcct
     4441 cccaggagac attatgaaag ttatgcgtac caccgtcgca actgttgtcg ccgcgacctt
     4501 atcgatgagt gctttctctg tgtttgcaga agcaagcctg acaggtgcag gtgcaacctt
     4561 ccctgcgccg gtgtatgcca aatgggctga cacttaccag aaagaaaccg gtaataaagt
     4621 taactaccag ggtatcggtt cttccggtgg cgtaaaacag attatcgcta ataccgttga
     4681 ttttggtgcc tctgacgcgc c
//
