LOCUS       sequence-316779-        6333 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Hydrocarbon Structures for Fuels and Chemicals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: JGI_018141
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     167..186
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        167..185
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    186..210
                     /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             225..247
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             266..283
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             293..310
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1364..1381
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1434..1452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1448..1495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1532..1987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1619..1638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1829..1850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2079..2894)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2079..2894)
                     /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     2802..2821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2886..2905
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3016..3604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3505..3524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3758..3775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3790..3929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3855..3874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4015..4037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4031..4222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5031..6113)
                     /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"
     primer_bind     6075..6094
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6114..6191)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
       61 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      121 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
      181 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
      241 aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg gcctggtgcc
      301 gcgcggcagc catatgaaat accgtgcggt caccttggaa agttttggat accagctcgc
      361 gccagttgtg gtgtctacct ctgacctgga agcgcgtttg gagccgcttt atcggcaatt
      421 gaggatcgcg ccagggcagc tgcaggcaat gacaggcatc cgcgaacggc gttggtggga
      481 accaggatat ccactttcac aaggcgcaat tgcggcagct cggaaagcgc ttgccgcctc
      541 ggcggttcct gttgaccaaa tcggggcgct tatatacgcg ggtgtttgcc gcgaacagtt
      601 tgaacctgcc actgcttgtc gggtcgcgca tggtttaggg atcggaggtc aagcggcagt
      661 ctttgatatc tctaacgcgt gcctgggcgt gctgaacggt atgttagatt tagctaaccg
      721 catcgaatta ggtcaaatac gggcaggtat ggtggtaagc tgcgaatcgg cacgtgaaat
      781 taacgaaatc atgattgaac gtatgctgaa ggaacgtcgc atggagcatt tcgcgagttc
      841 tctggccacc ttaacgggcg gtagtggtgc cgttgcggta ctacttacgg atgggtcttt
      901 tagcggcgca cagcgtcgta aactgttggg cggagtgtgc ctgtccgccc cggaacacca
      961 ccagttttgc ctttggggta tggcgccaga tggacgtggg ggttatcagc aggtgatgag
     1021 caccgatggc gttaacgtca tgaaccatgg cgttgaatta ggtaaacgca cgtggaacgc
     1081 gttcttacca catctgggtt ggtcagctga acaggtggat caggttatct gtcaccaagt
     1141 tggcagtgcg catcaaaatg ccatcctgaa gactttagga ataagtgccg accaagattt
     1201 cactacctac gagtacctgg gtaatatggg tacagtttct ttacctttaa cggctgcgtt
     1261 ggccgcggag agagacattc ttttgactgg agatcgcgta gccctgcttg ggatcggttc
     1321 gggtcttaat tgcttaatgc tgggggtaga atggtaactc gagcaccacc accaccacca
     1381 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1441 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1501 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1561 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1621 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1681 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1741 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1801 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1861 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1921 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1981 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2041 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     2101 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2161 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2221 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2281 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2341 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2401 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     2461 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     2521 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     2581 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     2641 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     2701 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     2761 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     2821 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     2881 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3121 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3781 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     3841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3901 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4081 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4141 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4201 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     4261 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     4321 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4381 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     4441 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     4501 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4561 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4621 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4681 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     4741 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     4801 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4861 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4921 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4981 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     5041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5101 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     5161 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     5221 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     5281 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     5341 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     5401 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     5461 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     5521 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5581 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     5641 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     5701 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     5761 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     5821 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5881 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5941 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     6001 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     6061 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     6121 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     6181 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     6241 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     6301 ggcgcccaac agtcccccgg ccacggggcc tgc
//
