Ppj220(JBEI-13483)

LOCUS       Ppj220(JBEI-13483)                6730 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  for expression of C-StrepII-3607.
ACCESSION   .
VERSION     .
KEYWORDS    Ppj220(JBEI-13483)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  AUTHORS   Javidpour P, Deutsch S, Mutalik VK, Hillson NJ, Petzold CJ, Keasling
            JD, Beller HR
  TITLE     Investigation of Proposed Ladderane Biosynthetic Genes from Anammox 
            Bacteria by Heterologous Expression in E. coli.
  JOURNAL   PLoS One. 2016 Mar 14;11(3):e0151087. doi: 
            10.1371/journal.pone.0151087. eCollection 2016.
  PUBMED    26975050
REFERENCE   2  (bases 1 to 6730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6730
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     921..940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(960..1037)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1243..1262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1346..1365
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1346..1364
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1365..1389
                     /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)."
     RBS             1404..1426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2907..2930
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2940..2957
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3010..3028)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3024..3071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3108..3563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3195..3214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3405..3426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3655..4470)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4378..4397
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4462..4481
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4592..5180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5081..5100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5334..5351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5366..5505
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5431..5450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5591..5613
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5607..5798)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6607..959)
                     /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"
ORIGIN
        1 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
       61 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      121 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
      181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
      481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
      541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
      601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
      661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
      721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
      781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
      841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
      901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
      961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     1021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     1081 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     1141 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     1201 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     1261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     1321 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     1381 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatgagta
     1441 gcaccaaacc aattaaacca catattcaaa gtaccaagat tgacacgctg tatccgagtt
     1501 acgatgtgat tgttattggt gccggtatcg ccggtctgat ttgtgccgcc ttcctggcca
     1561 aaagtggtaa gaaagccctg ctgattgagc aacatttcat tccaggtggc tactgtacca
     1621 gctttaaacg taagggtttc ctgtttgatg cggcggttca tcatattggc ggttgcggta
     1681 aatggagcgt ggtgggtcgt tgtttcaaaa tgctgggcat cgaaatggac ttttaccatc
     1741 tggacccaat ggataatctg atcttcccaa cgtttagcat tgaaattcca gccgatctgg
     1801 atgaatacct gctgcgcctg caaaagcgtt acccacatga aaaggataac attaagtggt
     1861 tttttcaaga ctttatccgc ctgtaccgcg ccaccttcca taatgagaaa agtcaaatca
     1921 tcgagaaata caaagatatg acgtacggca tgatgctgga tcatttcttt gatgatagtg
     1981 aactgaaaat ggttctgagc ggccagtggg gttacgtggg tctggccccg agcaaggcca
     2041 gcgccattgc catgagccaa atgatgatta actatctgaa agatggtgcc ttttttccaa
     2101 agggtggtac ccaagaattt gccaatgcca tttttcgtaa gtttatcgat ttcggcggtc
     2161 atgttatgct gagtagtaaa gtgaagaaaa tcctgaagaa gggcccaagt gttaatggtg
     2221 tgaaactgaa tgacgataag gaaattcata gcagtgttgt ggtgagcaat atcgatgcgc
     2281 gtcagacctt ttatcatctg ctgaacgacg gtaatgatgt ggatgccgtt tacctgaaaa
     2341 acattgaggg tatgaaagaa agttgtagtt tttttctgct gtacctgggt atcggtaaag
     2401 gcattgacct gaatacgctg aagcgcggct tctacctgag tggtagcaat ttcgacagca
     2461 tcgacagtga atggatgtat attagtgtgc caaccagtat ttgtcaagcg ctgagtccac
     2521 aaaataagca aattatcagt atcgtggtgt acaagaaaga agagttctac aaaaatgtga
     2581 ccgattggga actgctgaaa cagaatattg tgagcagtac gctggagcgc ctggaaagtt
     2641 acctgccgga tattaagaaa catattgaga tcattgaagt tgccacccca aaaaccctgg
     2701 aacgctacac cctgaacacg aacggtgccg cgtatggttg ggccgtgagt gttgaccaaa
     2761 tctgggaaca gcgtctgagc ccaacgacgc cgattagtgg tctgtatctg gcgggtcatt
     2821 ggacgcgccc aggtccaggt atctgcgccg tggttagtag tggttggagt gtggccaacc
     2881 tgattctgaa aaattggcgt agcgcttgga gccacccgca gttcgaaaaa taactcgagc
     2941 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3001 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     3061 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
     3121 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3181 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3301 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3421 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3481 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3541 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     3601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     3661 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     3721 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     3781 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     3841 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3901 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     3961 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     4021 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     4081 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     4141 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     4201 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     4261 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     4321 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     4381 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     4441 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     4501 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4561 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4621 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4681 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4741 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4801 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4861 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4921 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4981 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5041 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5101 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5161 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5221 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5281 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5341 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5401 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5461 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     5521 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5581 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5641 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     5701 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     5761 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     5821 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     5881 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     5941 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     6001 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     6061 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     6121 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     6181 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     6241 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     6301 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     6361 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     6421 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     6481 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     6541 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     6601 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6661 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     6721 cagtgagacg
//