LOCUS       sequence-316001-        6294 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 6294)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6294)
  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_018061
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             56..73
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             83..100
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1115..1132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1185..1203)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1199..1246
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1283..1738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1370..1389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1580..1601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1830..2645)
                     /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(1830..2645)
                     /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     2553..2572
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2637..2656
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2767..3355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3256..3275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3509..3526
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3541..3680
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3606..3625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3766..3788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3782..3973)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4782..5864)
                     /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     5826..5845
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5865..5942)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6148..6167
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6251..6270
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6251..6269
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6270..6294
                     /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)."
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat accatgggca gcagccatca
       61 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatggttc atcagaacgt
      121 agcacttctt gccctcgcga cgaccttacc cgatgatgtt actaccagtg atgagattga
      181 tgaacggctg ggggaaacac tgcgacgcct acgcgtccca cgtcgcatgt tacagcgcgt
      241 cgcgggcgtt catgaacgca gacgttgggc gaaggaccag agttttgtat ccgcagcggt
      301 tgcggccgga gaaaaagcac tggcggaagc gggcgtcgcc ccgtctgaag tgggtttgat
      361 tataaacact tcggtgacta ggccgcattt agagccgtct gtagcagtac ggatgcacga
      421 cggtttggga ctgccgtcca gtgcaatgaa tttcgatatt gcaaatgcgt gcctgggctt
      481 cgtaaatggg atgaccgttg cggcctcgat gatcgatgca ggtcagataa ggtatgccat
      541 cgttatcgat ggtgaagacg ccgatgaaat gcaggatcgg actattgaaa ggctgagccg
      601 tccgacaacc acgcgtgaag atttcatgaa tgaattcgct tcccttacct tgggtagcgg
      661 ggccgcggcc gtggtgctgg gacctgcaga tgcacaccca gtgggccatc gcattctggg
      721 tggagtgact cgtgccgcga cacagtttca tgaagtgtgc gttggatctg tagatggcat
      781 gatcactgat acgaaggcgc tgcttaaggg aggtatggaa ctggtcgtag cagcgtggaa
      841 ggatgcccgt gaacatttct catggagctc catggatagg tatatccttc atcaggttag
      901 tgatgttcat actgatgcaa ttgtcaaggc tgcccgtcta gatcgcagta aagttccact
      961 gacatatccg cgacacggaa acgtaggtcc cgcttcaata ccgatcacac tggctgaaca
     1021 ggctccgagc cttgcgcctg gggatagagt tcttctgatg ggcgttggca gcggtcttaa
     1081 tacgtcgatg atggaaattg cctggtaact cgagcaccac caccaccacc actgagatcc
     1141 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1201 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1261 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1561 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1621 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1681 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1741 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1801 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     1861 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1921 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1981 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2041 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     2101 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2161 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     2221 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2281 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     2341 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2401 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2461 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     2521 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2581 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     2641 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     2701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2881 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3361 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3421 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3481 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3541 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3601 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3661 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3841 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3901 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3961 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     4021 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     4081 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     4141 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     4201 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     4261 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4321 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4381 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4441 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4501 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4561 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4621 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4681 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4741 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4801 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4861 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4921 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4981 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     5041 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     5101 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     5161 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     5221 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5281 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5341 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5401 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5461 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5521 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5581 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5641 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5701 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5761 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5821 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5881 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5941 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     6001 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6061 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6121 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6181 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     6241 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcc
//
