LOCUS       SEGA028                4423 bp ds-DNA     linear   UNA 22-JUN-2021
DEFINITION  LMBP 12864.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4423)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: Ptrc-high-TEV-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4423
                     /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..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"
     misc_feature    1795..1824
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    1795..1800
                     /label=-35
     -35_signal      1795..1800
                     /gene="trp"
                     /label=trp
     misc_feature    1818..1824
                     /label=-10
     -10_signal      1818..1824
                     /gene="lacUV5"
                     /label=lacUV5
     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..1898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="MENLYFQ"
     CDS             1899..2609
                     /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
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2511..2513
                     /label=A206T
     misc_feature    2613..2661
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2672..2700
                     /label=Promoter 3BCD2
     CDS             2740..3930
                     /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    3911..3960
                     /label=HR3
     terminator      3941..3981
                     /label=rrnC
     misc_feature    3982..4031
                     /label=homology
     CDS             4176..4423
                     /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 aatatcgatg gagaacctgt acttccagag caaaggagaa gaacttttca
     1921 ctggagttgt gccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     1981 tcagtggaga gggtgaaggt gatgctacat acggaaaact cacccttaaa tttatttgca
     2041 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactctgacc tatggcgttc
     2101 aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc
     2161 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaac tacaagacgc
     2221 gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg
     2281 attttaaaga agatggaaac attctcggac acaaactaga gtacaactat aactcacaca
     2341 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc
     2401 acaacattga agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg
     2461 gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct accctttcga
     2521 aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga
     2581 ttacacatgg catggatgag ctctacaaat gaccaattat tgaaggccgc taacgcggcc
     2641 tttttttgtt tctggtctcc caaaaattta tttgcttatt aatcatccgg ctcgtataat
     2701 gtgtggaggg cccaagttca cttaaaaagg agaatagtta tgaaatcgaa caatgcgctc
     2761 atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta
     2821 ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg
     2881 ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc
     2941 gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac
     3001 gcgatcatgg cgaccacacc cgtcctgtgg attctctacg ccggacgcat cgtggccggc
     3061 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     3121 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     3181 cccgtggccg ggggactgtt gggcgccatc tccctgcacg caccattcct tgcggcggcg
     3241 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     3301 gagcgccgtc cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3361 ggcatgacca ttgtggccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3421 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     3481 agcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     3541 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
     3601 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
     3661 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
     3721 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
     3781 tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
     3841 gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
     3901 ggtgcatgga gccgggccac ctcgacctga gacaattgtc cagataaaaa aaatccttag
     3961 ctttcgctaa ggatgatttc taaaattttt ctgtcatctc ttcgttatta atgtttgtaa
     4021 ttgactgaat atcaacgctt atttaaatca gactgaagac tttatctctc tgtcataaaa
     4081 ctgtcatatt ccttacatat aactgtcacc tgtttgtcct attttgcttc tcgtagccaa
     4141 caaacaatgc tttatgaatc ctcccaggag acattatgaa agttatgcgt accaccgtcg
     4201 caactgttgt cgccgcgacc ttatcgatga gtgctttctc tgtgtttgca gaagcaagcc
     4261 tgacaggtgc aggtgcaacc ttccctgcgc cggtgtatgc caaatgggct gacacttacc
     4321 agaaagaaac cggtaataaa gttaactacc agggtatcgg ttcttccggt ggcgtaaaac
     4381 agattatcgc taataccgtt gattttggtg cctctgacgc gcc
//
