LOCUS       pJP_Ctrl10        6295 bp DNA     circular SYN 14-NOV-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6295)
  AUTHORS   UNIL
  TITLE     Direct Submission
  JOURNAL   Exported Apr 11, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     CDS             1..40
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /note="color: #993366"
                     /translation="TLENILAKYGRI"
     primer_bind     1..20
                     /label=pr_JP105
                     /note="color: black; sequence: cacactagagaacatactgg; added:
                     2023-10-10"
     primer_bind     complement(1..20)
                     /label=pr_JP108
                     /note="color: black; sequence: ccagtatgttctctagtgtg; added:
                     2023-10-10"
     primer_bind     559..581
                     /label=pr_JP111
                     /note="color: black; sequence: 
                     gtacctttctcctctttaatgaattctagtacatgcaaccattatcaccgccagaggta
                     aaatagtcaacacgcacggtgttagatatttatcctcgacctgcagctaggtctagggc
                     gg; added: 2023-10-18"
     terminator      585..671
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     CDS             complement(736..1815)
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(936..955)
                     /label=pr_JPseq33
                     /note="color: black; sequence: caccatcaaacaggattttc; added:
                     2023-10-10"
     primer_bind     1623..1642
                     /label=pr_JPseq31
                     /note="color: black; sequence: caatcagcaacgactgtttg; added:
                     2023-10-10"
     primer_bind     complement(1675..1693)
                     /label=pr_JPseq32
                     /note="color: black; sequence: gatggcggagctgaattac; added: 
                     2023-10-10"
     primer_bind     1827..1848
                     /label=Primer 1
                     /note="color: black; sequence: ggtacctttctcctctttaatg; 
                     added: 2023-11-14"
     primer_bind     complement(1827..1846)
                     /label=pr_JP110
                     /note="color: black; sequence: ttaaagaggagaaaggtacc; added:
                     2023-10-18"
     primer_bind     complement(1828..1947)
                     /label=pr_JP104
                     /note="color: black; sequence: 
                     ccgccctagacctagctgcaggtcgaggataaatatctaacaccgtgcgtgttgactat
                     tttacctctggcggtgataatggttgcatgtactagaattcattaaagaggagaaaggt
                     ac; added: 2023-10-10"
     primer_bind     1828..1947
                     /label=pr_JP111
                     /note="color: black; sequence: 
                     gtacctttctcctctttaatgaattctagtacatgcaaccattatcaccgccagaggta
                     aaatagtcaacacgcacggtgttagatatttatcctcgacctgcagctaggtctagggc
                     gg; added: 2023-10-18"
     RBS             1833..1844
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /note="color: #a6acb3"
     primer_bind     1931..1965
                     /label=pr_JP106
                     /note="color: black; sequence: 
                     agctaggtctagggcggAACGATTACCAATACTTC; added: 2023-10-10"
     misc_feature    1948..2138
                     /label=2018mar28_200bpSpacer2.5
                     /note="color: #ffcc99"
     primer_bind     2033..2052
                     /label=pr_JPseq16
                     /note="color: black; sequence: TCTGTTATCGCTACCCTGTC; added:
                     2023-03-11"
     misc_feature    2139..2199
                     /label=L3S2P21 Terminator
                     /note="color: #a6acb3"
     primer_bind     complement(2185..2203)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     gctagcactgtacctaggactgagctagccgtcaaAGGCTCGGTACCAAATTC; 
                     added: 2023-11-14"
     misc_feature    2203..2225
                     /label=PT3
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     2226..2247
                     /label=Primer 3
                     /note="color: black; sequence: 
                     ttgacggctagctcagtcctaggtacagtgctagcTTGTGAGCGGATAACAATtCGC; 
                     added: 2023-11-14"
     protein_bind    2226..2242
                     /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)."
                     /note="color: #31849b"
     protein_bind    2249..2265
                     /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)."
                     /note="color: #31849b"
     primer_bind     2282..2318
                     /label=pr_JP61
                     /note="color: black; sequence: 
                     gcaggacgcactgaccgtacagtgctagcTAATTTTG; added: 2023-03-01"
     RBS             2315..2337
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             2345..3055
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2590..2609)
                     /label=pr_EB002
                     /note="color: black; sequence: TCAAGTAGTCGGGGATGTCG; added:
                     2023-10-19"
     primer_bind     2897..2914
                     /label=pr_JPseq36
                     /note="color: black; sequence: ACCTACAAGGCCAAGAAG; added: 
                     2023-10-10"
     terminator      3230..3277
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     terminator      3286..3372
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff; direction: RIGHT"
     terminator      3464..3491
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(3484..3521)
                     /label=pr_JP62
                     /note="color: black; sequence: 
                     aaaagtgccacctgacgtATCTGAAACGCAAAAAGGCC; added: 2023-03-01"
     promoter        3534..3638
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(3639..3707,3708..4499)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3639 .. 3707 / #ccffcc / signal sequence
                        2: 3708 .. 4499 / #ccffcc
                     Cleavage site after base 3707"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3641..3660)
                     /label=pr_JPseq15
                     /note="color: black; sequence: cacggaaatgttgaatactc; added:
                     2023-03-11"
     primer_bind     complement(3907..3926)
                     /label=pr_JP107
                     /note="color: black; sequence: agaatagtgtatgcggcgac; added:
                     2023-10-10"
     primer_bind     3909..3927
                     /label=pr_JP109
                     /note="color: black; sequence: cgccgcatacactattctc; added: 
                     2023-10-10"
     primer_bind     complement(4116..4133)
                     /label=pr_JPseq35
                     /note="color: black; sequence: cggttcccaacgatcaag; added: 
                     2023-10-10"
     terminator      4529..4623
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
     rep_origin      5115..5337
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
                     /note="color: #ffff00"
     CDS             join(5385..6295,1..20)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /note="color: #993366"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENIL"
     primer_bind     6044..6063
                     /label=pr_JPseq34
                     /note="color: black; sequence: ccaagttgaactagatagac; added:
                     2023-10-10"
ORIGIN
        1 cacactagag aacatactgg ctaaatacgg aaggatctga ggttcttatg gctcttgtat
       61 ctatcagtga agcatcaaga ctaacaaaca aaagtagaac aactgttcac cgttacatat
      121 caaagggaaa actgtccata tgcacagatg aaaacggtgt aaaaaagata gatacatcag
      181 agcttttacg agtttttggt gcattcaaag ctgttcacca tgaacagatc gacaatgtaa
      241 cagatgaaca gcatgtaaca cctaatagaa caggtgaaac cagtaaaaca aagcaactag
      301 aacatgaaat tgaacacctg agacaacttg ttacagctca acagtcacac atagacagcc
      361 tgaaacaggc gatgctgctt atcgaatcaa agctgccgac aacacgggag ccagtgacgc
      421 ctcccgtggg gaaaaaatca tggcaattct ggaagaaata gcgctttcag ccggcaaacc
      481 ggctgaagcc ggatctgcga ttctgataac aaactagcaa caccagaaca gcccgtttgc
      541 gggcagcaaa acccgtacac ctgcagctag gtctagggcg gcggatttgt cctactcagg
      601 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
      661 cgttttattt gatgcctcta gactcagcta attaagcttt taagctgcta aagcgtagtt
      721 ttcgtcgttt gctgcctgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
      841 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
      901 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
      961 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     1021 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     1081 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     1141 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     1201 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     1261 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     1321 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     1381 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     1441 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     1501 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     1561 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     1621 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     1681 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     1741 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     1801 cgttactggt ttcacattca ccatatggta cctttctcct ctttaatgaa ttctagtaca
     1861 tgcaaccatt atcaccgcca gaggtaaaat agtcaacacg cacggtgtta gatatttatc
     1921 ctcgacctgc agctaggtct agggcggaac gattaccaat acttctgact ggatttcctg
     1981 agcggttctc ttcacgcaac tacgagcacc tactgtgtct tattacaatc tatctgttat
     2041 cgctaccctg tcccactgaa aagtctacgg aactgctgag tattattgat ttacctgcga
     2101 gaacgaacaa cactaccaga cctatccttt atctattagg accaaaacga aaaaaggccc
     2161 ccctttcggg aggcctcttt tctggaattt ggtaccgagc cttaataacc ctcactatag
     2221 ggagattgtg agcggataac aattcgcttt gtgagcggat aacaagatac tgagcacatc
     2281 agcaggacgc actgaccgta cagtgctagc taattttgtt taactttaag aaggagatat
     2341 acatatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     2401 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     2461 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     2521 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     2581 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     2641 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctctctgca
     2701 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2761 cgtaatgcag aagaagacta tgggctggga ggcctcctcc gagcggatgt accccgagga
     2821 cggcgcgctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2881 cgctgaagtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcgtacaa
     2941 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     3001 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaactag
     3061 ttgcggccgc ctgcgcacca cctgcaggac ctcagcaatc gcttggacca gctttcttaa
     3121 gggtacctat tacagctgga tccggcgccg ctcattcgct aatcgccacg acgcgtagtc
     3181 gacccgccaa ttcccatggg gcgcgcctgc tgccaccgct gagcaataac tagcataacc
     3241 ccttggggcc tctaaacggg tcttgagggg ttttttgcca ggcatcaaat aaaacgaaag
     3301 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3361 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
     3421 cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc catcctgacg
     3481 gatggccttt ttgcgtttca gatacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3541 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3601 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3661 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3721 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3781 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3841 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3901 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3961 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4021 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4081 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4141 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4201 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4261 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4321 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4381 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4441 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4501 tgtcagacca agtttacgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
     4561 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     4621 aatccaagca ctagggacag taagacgggt aagcctgttg atgataccgc tgccttactg
     4681 ggtgcattag ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa
     4741 atcagagggc aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc
     4801 cataaaacgc cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc
     4861 atgaatccat aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag
     4921 cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa
     4981 aaggaatatt cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt
     5041 aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa
     5101 ctgactaaag tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt
     5161 ctttctttat tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc
     5221 tagcggaatt tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga
     5281 atttacagat acccacaact caaaggaaaa ggactagtaa ttatcattga ctagcccatc
     5341 tcaattggta tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg
     5401 ttttcaaagc aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc
     5461 taattttatg ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac
     5521 ggacggtatc gttcacttat aaccaatacg ctcagatgat gaacatcagt agggaaaatg
     5581 cttatggtgt attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga
     5641 atcctttggt taaaggcttt gagattttcc agtggacaaa ctatgccaag ttctcaagcg
     5701 aaaaattaga attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat
     5761 tcataaaata taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt
     5821 atgagtggtt attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta
     5881 gccttgatga atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc
     5941 ttaaccaatg ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat
     6001 tggtggttga taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata
     6061 gacaaatgga tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa
     6121 taccaacaac cattacatca gattcctacc tacataacgg actaagaaaa acactacacg
     6181 atgctttaac tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc
     6241 aaagtaagta tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaac
//
