LOCUS       sequence-317616-        6330 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 6330)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6330)
  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_017981
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1195..1212
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1265..1283)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1279..1326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1363..1818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1450..1469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1660..1681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1910..2725)
                     /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(1910..2725)
                     /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     2633..2652
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2717..2736
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2847..3435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3336..3355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3589..3606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3621..3760
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3686..3705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3846..3868
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3862..4053)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4862..5944)
                     /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     5906..5925
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5945..6022)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6228..6247
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg gcttcaaccg cgacgaccag tctgcgcaac
      181 gcatatcgct tacaagatac tgtgattctg tctgccaccg cagttgaggc cccagaacgt
      241 gtcaccagcg cttggattga cgagcaactc agcgagacct attcacggct aagcatgacc
      301 ccgggcaagt ttgaggaact ggcaggtgtc gtggctcgcc gctggtggcc cagtggcgtc
      361 tcctttatag atggagcagc agaagcaggg gcaaaagctc tcgcagaagc cggcgtggaa
      421 ccgggacaag tggggcttct gattaatacc agcgtttcta aagatcacct tgaaccatct
      481 gcagctgtcc aagtacatga cgcgattggc ctgccggcta cctgtctgaa ttttgatatt
      541 tccaacgcct gtcttggctt tataaacgga atgcaggtgg ctggaactat gattgatgcg
      601 gggcagattg attacgccct gattgtgaac agtgagaata tacggggatt acaggagaaa
      661 gcgatcgcac gcctgcagcg tccggagacc acggttgcgg aagttcgtga gcagtttgcc
      721 acattaaccc tgggttgtgg ggcgaccgct atggttttgg gccggggaga ccaaagctcg
      781 gggggtcatc gcctcttagg gggtactcat aaaagtgcaa cgcatcatca tgatctctgt
      841 gttggcgatc tttcgtttat gagaaccgac acacgcggtc tgtttgaggc aggattagca
      901 ctggctctcg aaaccttcga agactccgcg gaagagtttg gatggcatga ggtcgaccac
      961 ttagtcagcc atcagacttc tctggtgcat atagctgcgc tatgcgctcg cctggggctg
     1021 gatcaggcga aatttcctct tagcctgcca gaatttggta atatgggccc agccgcggcg
     1081 ccgtttacat tagctaccag ccaggatcgt tttgaaccag gagatagagt tatgatgctc
     1141 ggaattggct caggtctcaa cacatcattc gccgatatcc tttggtgact cgagcaccac
     1201 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1261 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1321 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     1381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1561 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1801 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1861 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     1921 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1981 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2041 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2101 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2161 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2221 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2281 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2341 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2401 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2461 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2521 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2581 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2641 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     2701 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     2761 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2821 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2881 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2941 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3001 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3061 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3481 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3541 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3601 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3661 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3721 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3781 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3841 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3901 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3961 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     4021 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     4081 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     4141 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     4201 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     4261 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     4321 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     4381 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     4441 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4501 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     4561 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     4621 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     4681 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     4741 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4801 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4861 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4921 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4981 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     5041 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     5101 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     5161 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     5221 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     5281 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     5341 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     5401 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     5461 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5521 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5581 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5641 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5701 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5761 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5821 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5881 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5941 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     6001 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     6061 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     6121 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     6181 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     6241 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     6301 tagaggatcg agatctcgat cccgcgaaat
//
