LOCUS       sequence-316873-        6318 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 6318)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6318)
  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_018125
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             524..541
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(594..612)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      608..655
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      692..1147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(779..798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     989..1010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1239..2054)
                     /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(1239..2054)
                     /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     1962..1981
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2046..2065
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2176..2764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2665..2684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2918..2935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2950..3089
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3015..3034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3175..3197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3191..3382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4191..5273)
                     /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     5235..5254
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5274..5351)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5557..5576
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5660..5679
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5660..5678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5679..5703
                     /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             5718..5740
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5759..5776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5786..5803
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 gtccggcagt accatagatc gcgatgactt tatgtcagag tttgcaagcc tgacgctggg
       61 ttccggcgcc gctgcagcag ttattggacc ggcggatagg catccggaag gccataggat
      121 cttaggcggc gtgactcgtg cagcgaccga gtttaatcac ctgtgcgttg gcagtgtaga
      181 cggtatgttt acggatgcgc gtgcgttgct gaagggtggt ttagatcttg tcgtgtcagc
      241 ctggaaagaa gctgcggggg actcatggcg ttggtccaaa atggacagat atattaccca
      301 tcaggtgtcc agtattcaca cagactccat ggttaaggcg gcttccttgg acagaagccg
      361 cgttcccgtt acttatcctt atctgggtaa cgtcggtccg gcatccattc ctataacgct
      421 agcagatcag caaagcacgt tgaacaaagg tgatcgtgta cttctcatgg gtgtaggttc
      481 tgggttgaac acaggcctga tggagctggc ctggtgactc gagcaccacc accaccacca
      541 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      601 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      661 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
      721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1141 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1201 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     1261 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     1321 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1381 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1441 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     1501 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     1561 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     1621 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1681 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     1741 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     1801 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     1861 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     1921 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     1981 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     2041 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2281 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2821 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2881 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2941 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     3001 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3061 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3121 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3181 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3241 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3301 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3361 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3421 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3481 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3541 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     3601 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3661 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     3721 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     3781 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     3841 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     3901 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     3961 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4021 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4081 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4141 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4201 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4261 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4321 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4381 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4441 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     4501 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4561 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4621 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4681 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4741 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4801 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4861 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4921 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4981 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5041 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5101 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5161 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     5221 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     5281 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     5341 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5401 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     5461 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     5521 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     5581 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     5641 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     5701 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca
     5761 tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgg taaccggtaa
     5821 cgccaccacc agatttgata acgtagccat gttaagtgtg gcttcgacct taccgtcggc
     5881 ggtagtgacc tcggtggaaa ttgaggaagg cttggccgcc gctatgaagc gtgccaggct
     5941 gccaaagggt atattgcgac gtgtggcagg cgtcgtggag cgccgtcatt ggggaccaga
     6001 tgaaaactct gatgatgcta cagtttccgc aggccgtcgc gcacttgccg aggcaggtat
     6061 cagcccgtca caaatcgggc tgatgatcaa tacatcagtc acccgtcagc atcttgagcc
     6121 gtcggttgca gtgcgcatcc atgatggtct gggattacca tccagtgcta ttaactttga
     6181 tattgcaaat gcgtgcctgg ggtttgtcac agggatgacc atggcagctg cgatgattga
     6241 atcaggtcag atcgaatatg cattaattgt caacggtgaa gatgccagcg aggtccatgc
     6301 gaacaccatt gaacgcct
//
