LOCUS       sequence-316876-        6315 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 6315)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6315)
  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_018113
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             617..634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(687..705)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      701..748
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      785..1240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(872..891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1082..1103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1332..2147)
                     /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(1332..2147)
                     /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     2055..2074
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2139..2158
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2269..2857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2758..2777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3011..3028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3043..3182
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3108..3127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3268..3290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3284..3475)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4284..5366)
                     /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     5328..5347
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5367..5444)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5650..5669
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5753..5772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5753..5771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5772..5796
                     /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             5811..5833
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5852..5869
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5879..5896
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 cgcatcaatg atcgatgctg gcgctattga gtatgccttg gttgttgcag gagaagaccc
       61 gtctccttgg cacaaggctg cggtggaacg tctgcgagcc gacgatatta ctcgtgaaca
      121 ggtggttcag gagttcgcga ctctcacctt gggttgcggg gctgcggctg cagtggttgg
      181 gccggctgat aggcatccgg atggtcatcg aattgtgggc agcgtcaccc gtgcaggcac
      241 agaatttaat catctgtgca ttggtggcga cgacgggatg tttactgacg ctactggtct
      301 tctgaaccac ggtgtgggtc tggttgttga agcttggcat gaggcggagc aggatggatg
      361 ggactggcgg gaaatggata gctatgtaac tcatcaggtc agtaattctc acaccaataa
      421 gctgatagaa gatatgggat tagacgcaac taaggtacct ctaacctttc cgtactgggg
      481 gaatgtagca gccgccgccc tgccaatgac tctggccctg gaagcggaac gcctggaacc
      541 gggtgcacgg gttctttgcc tgggtgttgg gtctggcttg aatacaactc tgatggaaat
      601 tgcatggtaa ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
      661 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
      721 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
      781 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
      841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
      901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
      961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1201 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     1261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1321 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1381 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1441 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1501 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1561 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1621 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1681 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     1741 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1801 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     1861 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1921 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1981 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2041 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2101 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2161 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt
     2221 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2281 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2341 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2401 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2461 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2521 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2581 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2641 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2701 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2761 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2821 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2881 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2941 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3001 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3061 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     3121 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3181 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3241 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3301 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3361 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3421 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3481 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3541 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3601 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3661 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3721 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3781 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3841 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3901 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3961 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4021 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4081 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4141 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4201 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4261 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4321 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4381 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4441 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4501 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4561 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4621 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4681 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4741 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4801 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4861 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4921 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4981 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     5041 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     5101 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5161 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5221 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5281 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5341 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5401 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5461 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5521 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5581 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5641 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5701 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     5761 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     5821 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
     5881 ggtgccgcgc ggcagccata tgaccggaaa ctctaccatt actaaccgta atgtggcgat
     5941 actcagcgtg acaaaaactg aagccgatgt agttattccc agcacccgtt ttgatgagga
     6001 actcgccgaa acattccgtc gtctgagaat gccgaagggc ctgttggaac gtcttgcagg
     6061 tataaaggaa cgtagggggt gggcccccgg tcgccatttc actgatggag caactgaagc
     6121 cggtgaagcg gcactggccg aatccggcgt gagacctgag caaattggtt tgtttgtcaa
     6181 tgcatcagtg acgcgtgata attatgaacc tgctgtggcc gttggaatcc atgaacgtct
     6241 cggcctgccg tcgtcggctc tgaattttga tatgaccaat gcctgtctgg gtttcgtaaa
     6301 cgctctgacg gttgc
//
