LOCUS       SEGA034                4468 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12870.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4468)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: Ptrc-high-PelB-sfGFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4468
                     /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..1943
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             1944..2654
                     /codon_start=1
                     /label=sfGFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2658..2706
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2717..2745
                     /label=Promoter 3BCD2
     CDS             2785..3975
                     /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    3956..4005
                     /label=HR3
     terminator      3986..4026
                     /label=rrnC
     misc_feature    4027..4076
                     /label=homology
     CDS             4221..4468
                     /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 attaattatg aaatatctgc tgccgaccgc tgctgctggt ctgctgctcc
     1921 tcgctgccca gccggcgatg gcccgtaaag gcgaagagct gttcactggt gtcgtcccta
     1981 ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg
     2041 aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc
     2101 cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt
     2161 atccggacca catgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc
     2221 aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat
     2281 ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg
     2341 gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg
     2401 ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg
     2461 gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc
     2521 tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga
     2581 aacgcgatca catggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg
     2641 atgaactgta caaatgacca attattgaag gccgctaacg cggccttttt ttgtttctgg
     2701 tctcccaaaa atttatttgc ttattaatca tccggctcgt ataatgtgtg gagggcccaa
     2761 gttcacttaa aaaggagaat agttatgaaa tcgaacaatg cgctcatcgt catcctcggc
     2821 accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg
     2881 cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat
     2941 gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc
     3001 cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc
     3061 acacccgtcc tgtggattct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     3121 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     3181 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     3241 ctgttgggcg ccatctccct gcacgcacca ttccttgcgg cggcggtgct caacggcctc
     3301 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg ccgtccgatg
     3361 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gaccattgtg
     3421 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3481 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgagcgg cctgtcgctt
     3541 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     3601 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     3661 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     3721 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     3781 cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat cactggaccg
     3841 ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt
     3901 gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg
     3961 gccacctcga cctgagacaa ttgtccagat aaaaaaaatc cttagctttc gctaaggatg
     4021 atttctaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     4081 cgcttattta aatcagactg aagactttat ctctctgtca taaaactgtc atattcctta
     4141 catataactg tcacctgttt gtcctatttt gcttctcgta gccaacaaac aatgctttat
     4201 gaatcctccc aggagacatt atgaaagtta tgcgtaccac cgtcgcaact gttgtcgccg
     4261 cgaccttatc gatgagtgct ttctctgtgt ttgcagaagc aagcctgaca ggtgcaggtg
     4321 caaccttccc tgcgccggtg tatgccaaat gggctgacac ttaccagaaa gaaaccggta
     4381 ataaagttaa ctaccagggt atcggttctt ccggtggcgt aaaacagatt atcgctaata
     4441 ccgttgattt tggtgcctct gacgcgcc
//
