LOCUS       SEGA031                4459 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12867.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4459)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: Ptrc-high-PROTi-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4459
                     /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..1898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="MENLYFQ"
     misc_feature    1899..1934
                     /label=PROTi-Linker
     CDS             1935..2645
                     /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    2649..2697
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2708..2736
                     /label=Promoter 3BCD2
     CDS             2776..3966
                     /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    3947..3996
                     /label=HR3
     terminator      3977..4017
                     /label=rrnC
     misc_feature    4018..4067
                     /label=homology
     CDS             4212..4459
                     /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 acttccagtt tcgtagcaaa ggagaagaac
     1921 ttgtcactgg agttagcaaa ggagaagaac ttttcactgg agttgtgcca attcttgttg
     1981 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
     2041 ctacatacgg aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccat
     2101 ggccaacact tgtcactact ctgacctatg gcgttcaatg cttttcccgt tatccggatc
     2161 acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca
     2221 ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg
     2281 atacccttgt taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc
     2341 tcggacacaa actagagtac aactataact cacacaatgt atacatcacg gcagacaaac
     2401 aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggttccgttc
     2461 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     2521 acaaccatta cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc
     2581 acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct
     2641 acaaatgacc aattattgaa ggccgctaac gcggcctttt tttgtttctg gtctcccaaa
     2701 aatttatttg cttattaatc atccggctcg tataatgtgt ggagggccca agttcactta
     2761 aaaaggagaa tagttatgaa atcgaacaat gcgctcatcg tcatcctcgg caccgtcacc
     2821 ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc
     2881 gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg
     2941 caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc
     3001 ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc
     3061 ctgtggattc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     3121 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     3181 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     3241 gccatctccc tgcacgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     3301 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gccgtccgat gcccttgaga
     3361 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgaccattgt ggccgcactt
     3421 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     3481 ttcggcgagg accgctttcg ctggagcgcg acgatgagcg gcctgtcgct tgcggtattc
     3541 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     3601 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     3661 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     3721 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     3781 cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc
     3841 acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc
     3901 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     3961 acctgagaca attgtccaga taaaaaaaat ccttagcttt cgctaaggat gatttctaaa
     4021 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     4081 aaatcagact gaagacttta tctctctgtc ataaaactgt catattcctt acatataact
     4141 gtcacctgtt tgtcctattt tgcttctcgt agccaacaaa caatgcttta tgaatcctcc
     4201 caggagacat tatgaaagtt atgcgtacca ccgtcgcaac tgttgtcgcc gcgaccttat
     4261 cgatgagtgc tttctctgtg tttgcagaag caagcctgac aggtgcaggt gcaaccttcc
     4321 ctgcgccggt gtatgccaaa tgggctgaca cttaccagaa agaaaccggt aataaagtta
     4381 actaccaggg tatcggttct tccggtggcg taaaacagat tatcgctaat accgttgatt
     4441 ttggtgcctc tgacgcgcc
//
