LOCUS       pACB108_pET21b_y        6211 bp DNA     circular SYN 04-NOV-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6211)
  AUTHORS   Alissa Bleem
  TITLE     Direct Submission
  JOURNAL   Exported May 30, 2024 from SnapGene 7.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6211
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     1..40
                     /label=oACB446
     primer_bind     complement(1..38)
                     /label=oACB422
     CDS             19..36
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     19..36
                     /label=oACB151
     primer_bind     50..70
                     /label=oIS011
     terminator      103..150
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      187..642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(193..211)
                     /label=oACB049
     promoter        669..773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(774..842,843..1634)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1805..2393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2579..2721
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2823..3014)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3823..4905)
                     /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(4906..4983)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5189..5208
                     /label=oACB050
     promoter        5292..5310
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5311..5335
                     /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)."
     primer_bind     complement(5336..5395)
                     /label=oACB447
     primer_bind     complement(5342..5377)
                     /label=oIS012
     RBS             5350..5372
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     5358..5402
                     /label=oACB421
     misc_feature    5377
                     /label=UST
     gene            join(5378..6211,1..18)
                     /gene="fghA"
                     /locus_tag="PP_RS08335"
                     /old_locus_tag="PP_1617"
                     /label=fghA
     CDS             join(5378..6211,1..18)
                     /codon_start=1
                     /transl_table=11
                     /gene="fghA"
                     /locus_tag="PP_RS08335"
                     /old_locus_tag="PP_1617"
                     /product="S-formylglutathione hydrolase
                     PP_RS08335"
                     /EC_number="3.1.2.12"
                     /label=PP_RS08335 (yeiG)
                     /note="Derived by automated computational analysis 
                     usinggene prediction method: Protein Homology."
                     /db_xref="GI:489345198"
                     /inference="COORDINATES:same species:RefSeq"
                     /protein_id="WP_003252340.1"
                     /translation="MSLDNISCQKSFGGWHKRYRHHSKVLGCDMVFAVYLPPQAEQGEK
                     LPVLYWLSGLTCTDENFMQKAGAQRLAAELGLIIVAPDTSPRGEQVPGDPDGAWDFGLG
                     AGFYLNATQQPWAQHYRMHDYVVEELPALIEAHFPASGERSISGHSMGGHGALVCALRN
                     PGRYRSVSAFSPISNPMDCPWGEKAFSRYLGEDRGRWREWDASVLLAETPAGQCPPLLV
                     DQGDRDDFLEKQLKPEALEQAARKGGHAMTLRMQPGYDHSYYFIASFIEEHLRHHAVAL
                     GRV"
ORIGIN
        1 gtggcactgg ggcgggttca ccaccaccac caccactgag atccggctgc taacaaagcc
       61 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      121 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
      181 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      601 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcaggtggc acttttcggg
      661 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      721 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      781 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      841 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      901 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      961 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1021 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1081 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1141 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1201 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1261 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     1321 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1381 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1441 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1501 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1561 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1621 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1681 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1741 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1801 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1861 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1921 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1981 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2041 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2101 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2161 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2221 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2281 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2341 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2401 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2461 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2521 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2581 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tatggtgcac
     2641 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     2701 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2761 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2821 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     2881 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     2941 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     3001 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     3061 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     3121 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     3181 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     3241 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     3301 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     3361 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     3421 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     3481 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     3541 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     3601 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     3661 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     3721 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     3781 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     3841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     3961 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     4021 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     4081 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     4141 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     4201 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     4261 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     4321 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     4381 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     4441 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     4501 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     4561 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     4621 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     4681 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     4741 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     4801 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     4861 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     4921 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     4981 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     5041 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     5101 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5161 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     5221 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     5281 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     5341 agaaataatt ttgtttaact ttaagaagga gatatacatg agcctggata acatctcctg
     5401 ccagaagagc ttcggcggct ggcacaagcg ttaccggcat cattccaagg tgctgggttg
     5461 cgatatggtg ttcgctgtgt acctgccgcc gcaggccgag cagggcgaaa agctgccggt
     5521 gctgtactgg ctcagcggcc tcacctgcac cgacgagaac ttcatgcaga aggccggcgc
     5581 ccagcgtctg gccgccgagc tggggctgat catcgtcgcc cccgatacca gcccgcgtgg
     5641 cgagcaggtg ccgggcgatc cggacggcgc ctgggacttc ggcctgggcg ctggcttcta
     5701 cctcaacgcc acccagcaac cctgggccca gcactaccgc atgcatgact atgtggtgga
     5761 ggagttgcca gccctgatcg aggcacactt cccggcgtcg ggcgagcgca gcatcagcgg
     5821 ccactccatg ggtgggcatg gcgcgctggt gtgcgccttg cgcaatccgg ggcgctaccg
     5881 ctcggtgtcg gcattctcgc cgatcagcaa tccgatggat tgtccgtggg gcgagaaggc
     5941 cttcagccgc tacctgggtg aagaccgtgg gcgctggcgt gagtgggatg ccagcgtgct
     6001 gctggccgaa accccggcag gccaatgccc gccgttgctg gtggatcagg gcgaccgtga
     6061 cgacttcctc gagaagcaac tcaagcctga agcgctggaa caggcggcac gcaaaggtgg
     6121 ccatgcaatg accctgcgta tgcagccagg ctatgaccat agctactact tcattgccag
     6181 cttcatcgag gagcacctgc gccatcatgc g
//
