LOCUS       pSLS.540                6615 bp    DNA     circular UNA 18-JAN-2022
DEFINITION  pET-21(+) Eco1 RT and recombineering ncRNA, fabH G954A, a1/a2
            length: 22.
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
FEATURES             Location/Qualifiers
     rep_origin      12..467
                     /direction="LEFT"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        494..598
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             599..1459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     rep_origin      1630..2218
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
                     /label="rop"
     CDS             complement(3648..4730)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
                     /label="lacI"
     promoter        complement(4731..4808)
                     /gene="lacI"
     promoter        5117..5135
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    complement(5136..5160)
                     /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)."
                     /label="lac operator"
     RBS             complement(5202..5207)
                     /note="ribosome binding site"
     gene            5216..6178
                     /created_by="User"
                     /label="ec86RT"
     misc_feature    6194..6203
                     /created_by="Seth"
                     /modified_by="Seth"
                     /label="extended a1"
     misc_RNA        6204..6284
                     /created_by="Seth"
                     /label="msr"
     primer          6204..6215
                     /created_by="User"
                     /modified_by="Seth"
                     /label="a1"
     msd             complement(6274..6419)
                     /created_by="Max_Schubert"
                     /modified_by="Seth"
     misc_feature    6274..6290
                     /created_by="Seth"
                     /label="Flanking"
     misc_feature    6291..6302
                     /created_by="Seth"
                     /label="stem base"
     primer_bind     complement(6303..6392)
                     /Sequence="CCAGGGAACACAAATGCAAATTGCGTCATGTTTTAATCCTTATCTTA
                     GAAACGAACCAGCGCGGAGCCCCAGGTGAATCCACCGCCAAAG"
                     /Tm="82.1"
                     /%GC="48.9"
                     /modified_by="Seth"
                     /label="fabH_editor"
     misc_feature    6304..6392
                     /Original_Bases="CGATTTCCGCAGCAGTGAAAGAGTTCTTCGGTTCCAGCCAG
                     CTGCCTCAGTTTATGGACCAGAACAACCCGCTGTCTGAGATTACGCAC"
                     /label="CGATTTCCGCAGCAGTGAAAGAGTTCTTCGGTTCCAGCCAGCTGCCTCAG
                     TTTATGGACCAGAACAACCCGCTGTCTGAGATTACGCAC"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(6393..6404)
                     /created_by="Seth"
                     /label="stem base"
     misc_feature    complement(6405..6419)
                     /created_by="Seth"
                     /label="Flanking"
     primer          6421..6432
                     /created_by="User"
                     /modified_by="Seth"
                     /label="a2"
     misc_feature    6433..6442
                     /created_by="User"
                     /modified_by="Seth"
                     /label="extended a2"
     CDS             6459..6476
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /translation="HHHHHH"
                     /label="6xHis"
     terminator      complement(6543..6590)
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /label="T7 terminator"
ORIGIN      
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     5161 cctctaggat ccgaattcaa ataattttgt ttaactttaa gaaggagata tacatatgaa
     5221 atccgctgaa tatttgaaca cttttagatt gagaaatctc ggcctacctg tcatgaacaa
     5281 tttgcatgac atgtctaagg cgactcgcat atctgttgaa acacttcggt tgttaatcta
     5341 tacagctgat tttcgctata ggatctacac tgtagaaaag aaaggcccag agaagagaat
     5401 gagaaccatt taccaacctt ctcgagaact taaagcctta caaggatggg ttctacgtaa
     5461 cattttagat aaactgtcgt catctccttt ttctattgga tttgaaaagc accaatctat
     5521 tttgaataat gctaccccgc atattggggc aaactttata ctgaatattg atttggagga
     5581 ttttttccca agtttaactg ctaacaaagt ttttggagtg ttccattctc ttggttataa
     5641 tcgactaata tcttcagttt tgacaaaaat atgttgttat aaaaatctgc taccacaagg
     5701 tgctccatca tcacctaaat tagctaatct aatatgttct aaacttgatt atcgtattca
     5761 gggttatgca ggtagtcggg gcttgatata tacgagatat gccgatgatc tcaccttatc
     5821 tgcacagtct atgaaaaagg ttgttaaagc acgtgatttt ttattttcta taatcccaag
     5881 tgaaggattg gttattaact caaaaaaaac ttgtattagt gggcctcgta gtcagaggaa
     5941 agttacaggt ttagttattt cacaagagaa agttgggata ggtagagaaa aatataaaga
     6001 aattagagca aagatacatc atatattttg cggtaagtct tctgagatag aacacgttag
     6061 gggatggttg tcatttattt taagtgtgga ttcaaaaagc cataggagat taataactta
     6121 tattagcaaa ttagaaaaaa aatatggaaa gaacccttta aataaagcga agacctaaca
     6181 tttaactatg ataagattcc gtatgcgcac ccttagcgag aggtttatca ttaaggtcaa
     6241 cctctggatg ttgtttcggc atcctgcatt gaatctgagt tactgtctgt tttcctgaat
     6301 tcctttggcg gtggattcac ctggggctcc gcgctggttc gtttctaaga taaggattaa
     6361 aacatgacgc aatttgcatt tgtgttccct gggaattcag gaaacccgtt ttttctgacg
     6421 taagggtgcg catacggaat ctagcggccg cactcgagca ccaccaccac caccactgag
     6481 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     6541 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     6601 gaactatatc cggat
//
