LOCUS       sequence-316781-        6321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hydrocarbon Structures for Fuels and Chemicals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6321)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6321)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: JGI_018135
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(58..135)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     341..360
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     444..463
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        444..462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    463..487
                     /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             502..524
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             543..560
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             570..587
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1629..1646
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1699..1717)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1713..1760
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1797..2252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1884..1903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2094..2115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2344..3159)
                     /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(2344..3159)
                     /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     3067..3086
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3151..3170
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3281..3869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3770..3789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4023..4040
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4055..4194
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4120..4139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4280..4302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4296..4487)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5296..6321,1..57))
                     /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 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
       61 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
      121 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
      181 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
      241 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
      301 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
      361 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
      421 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
      481 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
      541 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atgaaatata
      601 ccaaagttgc gatccattcg tttgattatc ttgagccgga cgaattcttg acttcagatg
      661 acattgaaac gaaactggct ccagtgtata aacgcttgaa cctgccagcc ggcagattag
      721 aacttatgac tggaataaaa tctcgcggcc tttgggcgaa tggaatgaga ccgagtgaac
      781 tgtccacacg tgcggcggaa cgtgttcttg aaaagtcggg catagaccga aatgagatcg
      841 gtctgctgat acacgcttcg gtctgccgcg attttctgga acccagtaca gccagtgtgg
      901 tccatcacaa tctggaactt aatgaagatg cgatgattta tgacttgagt aatgcatgtt
      961 tgggcgtggt aaacgctatg gccatggcgt ctaatatgat tgaactgggt caaatcaaat
     1021 atgccctgat tgtttccggc gaaaacggcg gtccgctgtt aacccagact attgaacact
     1081 taaacaatga cgagactctg acaaggaaat ccattaaaaa atacatcgca aatctcacga
     1141 ttggtagcgc tgctgttgca tatcttctct gtgattcttc gttggctccg gacgccccgc
     1201 gtatattagg gggtgctgtt tcgaccgaca gttcggcaaa tattctgtgt caaggcgacg
     1261 gagatacaac ctccctgatg atggagacag acagtgagga gcttttgaaa cacggtttgg
     1321 aacttgccaa gaaaaattgg aataaaacca aacaagaact ttcattagaa aacaaagata
     1381 tcgactgggt ggtcggacat caagttggta cggctcatga aacgttgacg atgaaagcga
     1441 tggaactgca ggatcacaaa actttcacca cgtatgagac acttggtaat actggatcgg
     1501 cggccctgcc gatcacgctg gcgaagcttt ctgaaaaaca gaagatcaag aaaggcgaga
     1561 aagttgcttt attgggcata ggcagcggtc tgacgtcaat aatgctgggt gtccagtggt
     1621 aactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1681 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1741 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     1801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1861 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1981 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2161 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2221 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     2281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     2341 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2401 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2461 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2521 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2581 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2641 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2701 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     2761 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     2821 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     2881 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2941 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3001 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3061 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3121 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3181 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3421 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3901 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3961 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4021 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4081 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4141 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4201 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4261 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4321 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4381 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4441 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4501 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4561 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4621 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4681 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4741 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4801 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4861 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4921 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     4981 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     5041 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5101 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5161 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5221 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5401 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5461 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5521 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5581 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5641 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5701 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5761 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5821 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5881 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5941 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     6001 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     6061 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     6121 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     6181 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6241 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6301 ctggttcacc acgcgggaaa c
//
