LOCUS       pALS2_Ma-tRNApyl        5513 bp DNA     circular SYN 30-SEP-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pALS2 Ma GFP fluorescent reporter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  AUTHORS   Savanna's PC
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2023 from SnapGene 6.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..95)
                     /label=pALS2 Ma seq rev
                     /note="color: black; sequence: ccatacccacgccgaaacaag; 
                     added: 2020-08-14"
     promoter        373..397
                     /label=Lpp promoter
                     /label=Ipp p
                     /note="color: #ff58aa"
     gene            412..486
                     /label=Ma Pyl tRNA(6)
                     /label=Mb PylTwt (NC_007355.1)
                     /note="color: #15af04; direction: RIGHT"
     primer_bind     complement(428..479)
                     /label=Green1rev
                     /note="color: black; sequence: 
                     ccggggcgtcgaaccccgctggctaggttttagagacccgctggtcgccgg; added:
                     2020-09-25"
     primer_bind     447..503
                     /label=Green2for
                     /note="color: black; sequence: 
                     taaaacctagccagcggggttcgacgccccggtctctcgccaactagtatccttag; 
                     added: 2020-09-25"
     terminator      496..524
                     /label=rrnC term
                     /note="color: #ff58aa"
     primer_bind     complement(781..810)
                     /label=Green2rev
                     /note="color: black; sequence: 
                     gattactttctgttcgacttaagcattatg; added: 2020-09-25"
     primer_bind     complement(1047..1075)
                     /label=pACYC_p15Cm_pBad_terminator_rev
                     /note="color: black; sequence: 
                     GTGTGACCGTGTGCTTCTCAAATGCCTGAAGAGTTTGTAGAAACGCAAAAAGGCCATC"
     gene            1209..2070
                     /label=SpR
                     /note="color: #15af04; direction: RIGHT"
     primer_bind     1274..1283
                     /label=araC-Ma p15 for
                     /note="color: black; sequence: 
                     TCATGTTAGTCATGCCCCGCGCCCAGGATCTCACCGTCATCACCGAAACGCGCGAGGC;
                      added: 2020-08-14"
     primer_bind     1274..1283
                     /label=pACYC_p15Cm_pBad_arac_for
                     /note="color: black; sequence: 
                     CCGGAAGGAGCTGACTGGGTTGAAGGCTCTCATCACCGTCATCACCGAAACGCGCGAGG
                     C"
     primer_bind     2071..2130
                     /label=ORI swap for
                     /note="color: black; sequence: 
                     tgtatttagaaaaataaacaaataGCTAGCCGCTTCCTCGCTCACTGACTCGCTACGCT
                     C; added: 2020-07-10"
     rep_origin      complement(2241..2785)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
                     /note="color: #ffff00"
     primer_bind     2799..2818
                     /label=pALS2 Ma seq for
                     /note="color: black; sequence: TTAATCAGATAAAATATTTC; added:
                     2020-08-14"
     primer_bind     complement(2886..2920)
                     /label=ORI swap rev
                     /note="color: black; sequence: 
                     gcgggacaggcTGACAActcccgaGGATCCTGGGCGCGGGGCATGACTAACATGAGAAT
                     T; added: 2020-07-10"
     primer_bind     2891..2948
                     /label=araC-Ma p15 for
                     /note="color: black; sequence: 
                     TCATGTTAGTCATGCCCCGCGCCCAGGATCTCACCGTCATCACCGAAACGCGCGAGGC;
                      added: 2020-08-14"
     primer_bind     2921..2948
                     /label=pACYC_p15Cm_pBad_arac_for
                     /note="color: black; sequence: 
                     CCGGAAGGAGCTGACTGGGTTGAAGGCTCTCATCACCGTCATCACCGAAACGCGCGAGG
                     C"
     CDS             complement(3073..3951)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /note="color: #993366"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3978..4262
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     4264..4293
                     /label=sfGFP for
                     /note="color: black; sequence: 
                     CCCGTTTTTTGGGCTAACAGGAGGAATTAA; added: 2020-08-14"
     primer_bind     complement(4264..4293)
                     /label=araC rev
                     /note="color: black; sequence: 
                     TTAATTCCTCCTGTTAGCCCAAAAAACGGG; added: 2020-08-14"
     CDS             4296..5039
                     /codon_start=1
                     /label=sfGFP 150TAG
                     /note="color: #00ff00"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSH*VYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYKGSHHHHHH"
     CDS             5019..5036
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      5285..5371
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     5388..5407
                     /label=Green Ma tRNA seq
                     /note="color: black; sequence: TGAACGTTGCGAAGCAACGG; added:
                     2020-09-25"
     terminator      5463..5490
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     complement(join(5479..5513,1..25))
                     /label=sfGFP-Ma p15 rev
                     /note="gtcgacacagatgtaggtgttccacagggtAGAGTTTGTAGAAACGCAAAAA
                     GGCCATCC"
                     /note="color: black; sequence: 
                     accctgtggaacacctacatctgtgtcgacAGAGTTTGTAGAAACGCAAAAAGGCCATC
                     C; added: 2020-08-14"
     primer_bind     5479..5508
                     /label=Green1for
                     /note="color: black; sequence: 
                     GGATGGCCTTTTTGCGTTTCTACAAACTCT; added: 2020-09-25"
     primer_bind     complement(5480..5508)
                     /label=pACYC_p15Cm_pBad_terminator_rev
                     /note="color: black; sequence: 
                     GTGTGACCGTGTGCTTCTCAAATGCCTGAAGAGTTTGTAGAAACGCAAAAAGGCCATC"
ORIGIN
        1 cacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
       61 aatggtgcag ggcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
      121 gttgggcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
      181 ggcgacgata gtcatgcccc gcgcccaccg gaaggagcta ccggcagcgg tgcggactgt
      241 tgtaactcag aataagaaat gaggccgctc atggcgttct gttgcccgtc tcactggtga
      301 aaagaaaaac aaccctggcg ccgcttcttt gagcgaacga tcaaaaataa gtggcgcccc
      361 atcaaaaaaa tattctcaac ataaaaaact ttgtgtaata cttgtaacgc tagatctggg
      421 ggacggtccg gcgaccagcg ggtctctaaa acctagcata gcggggttcg acaccccggt
      481 ctctcgccaa ctagtatcct tagcgaaagc taaggatttt ttttaagctt ggcactggcc
      541 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      601 gcacatcccc ctttcgccag acgctctccc ttatgcgact cctgcattag gaagcagccc
      661 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagccc
      721 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctggcggccg
      781 cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta
      841 aattggatat cggcaggcca cgcgatcgct gacgtcggta ccctcgagtc tggtaaagaa
      901 accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta
      961 acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1021 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt
     1081 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     1141 cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt
     1201 gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     1261 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     1321 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     1381 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     1441 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     1501 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     1561 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     1621 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     1681 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     1741 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     1801 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     1861 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     1921 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     1981 gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc
     2101 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     2161 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     2221 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     2281 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2341 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     2401 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     2461 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     2521 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     2581 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     2641 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     2701 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     2761 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta
     2821 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     2881 gttgtaattc tcatgttagt catgccccgc gcccaggatc tcaccgtcat caccgaaacg
     2941 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
     3001 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     3061 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     3121 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     3181 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     3241 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     3301 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     3361 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     3421 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     3481 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     3541 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     3601 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     3661 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     3721 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     3781 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     3841 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     3901 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     3961 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     4021 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     4081 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     4141 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     4201 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     4261 atacccgttt tttgggctaa caggaggaat taaccatggt tagcaaaggt gaagaactgt
     4321 ttaccggcgt tgtgccgatt ctggtggaac tggatggtga tgtgaatggc cataaattta
     4381 gcgttcgtgg cgaaggcgaa ggtgatgcga ccaacggtaa actgaccctg aaatttattt
     4441 gcaccaccgg taaactgccg gttccgtggc cgaccctggt gaccaccctg acctatggcg
     4501 ttcagtgctt tagccgctat ccggatcata tgaaacgcca tgatttcttt aaaagcgcga
     4561 tgccggaagg ctatgtgcag gaacgtacca ttagcttcaa agatgatggc acctataaaa
     4621 cccgtgcgga agttaaattt gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggta
     4681 ttgattttaa agaagatggc aacattctgg gtcataaact ggaatataat ttcaacagcc
     4741 attaggtgta tattaccgcc gataaacaga aaaatggcat caaagcgaac tttaaaatcc
     4801 gtcacaacgt ggaagatggt agcgtgcagc tggcggatca ttatcagcag aataccccga
     4861 ttggtgatgg cccggtgctg ctgccggata atcattatct gagcacccag agcgttctga
     4921 gcaaagatcc gaatgaaaaa cgtgatcata tggtgctgct ggaatttgtt accgccgcgg
     4981 gcattaccca cggtatggat gaactgtata aaggcagcca ccatcatcat caccattaaa
     5041 gctcgagatc tgcagctggt accatatggg aattcgaagc ttggctgttt tggcggatga
     5101 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     5161 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     5221 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     5281 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     5341 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     5401 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     5461 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactctgt cga
//
