LOCUS       sequence-316874-        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_018119
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..37
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1073..1090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1143..1161)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1157..1204
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1241..1696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1328..1347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1538..1559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1788..2603)
                     /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(1788..2603)
                     /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     2511..2530
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2595..2614
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2725..3313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3214..3233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3467..3484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3499..3638
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3564..3583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3724..3746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3740..3931)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4740..5822)
                     /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     5784..5803
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5823..5900)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6106..6125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6209..6228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6209..6227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6228..6252
                     /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             6267..6289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(6308..6315,1..10)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tcatcatcac agcagcggcc tggtgccgcg cggcagccat atgaccaatt caaatgccac
       61 cttccgtcac caaaattcag cactgttagc agtgaaggcc agactggcgc ccctagtcgt
      121 gacgtctgaa gaaattgacg tacgtctcgc cgacgccctg accagattgc gcctcccgac
      181 aggcctgtta caaagagttg ccggggtgat ttcccgccgt aactgggacg ataccactac
      241 gtttgacgaa gccgctgccg gggcagcttc cgaagctctt aaggaggcta acgttgatcc
      301 ccaggatgtc gggttattaa tcaacaccag cgtgaccagg aaacatttgg agccttcagt
      361 agcagtcggc attcatcatg cactgggcct gccgagctct gccatgaatt ttgatatcac
      421 caatgcctgc ctgggtttcg tgaacggtat gactctggct tcccagatga ttgattccgg
      481 ccagataaaa tacgcggtta ttgtggatgg ggaggatagc aatgaaattc aggcgaacac
      541 tattgcacgg ctgtcacagc cgggaatctc ccggaaagac tttgtgaacg aatttgcctc
      601 acttactctg gggagcggcg ctgctgcagc agtcctgggg cctgccgacg cacatccgga
      661 gggacatcgg ataatgggag ggataagccg ggcaggaacc gaacatcatt tgctgtgcgt
      721 gggagataag gacggaatgt ttactcacac taaggaatta ctggcaggtg gcatggcgtt
      781 agttgtggaa gcgtggaagg aggcgaaggg cagttgggat tggcagaaaa tggataggta
      841 tattttgcat caagtttccg acgtacacac cgactctatc gtaaaagccg ctaagctgga
      901 ccgtaaacgt ataccactca cctacccttc tctagggaat gtggggccgg ctagtctacc
      961 catgaccctc gccctggagg caagcaagct ggtgaaggga aaccgagtgt tgtgcatggg
     1021 tgtcgggagc ggtatcaaca ccgccatgac agaaatagtt tggtgactcg agcaccacca
     1081 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1141 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1201 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1261 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1321 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1381 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1441 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1501 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1561 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1621 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1681 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1741 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     1801 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1861 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1921 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1981 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2041 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2101 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2161 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2221 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     2281 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     2341 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     2401 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     2461 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     2521 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     2581 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     2641 tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3481 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3541 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3601 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3661 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3721 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3781 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3841 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3901 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3961 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     4021 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     4081 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     4141 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     4201 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     4261 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     4321 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     4381 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     4441 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4501 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4561 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4621 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4681 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4741 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4801 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4861 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     4921 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4981 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     5041 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5101 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5161 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5221 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5281 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5341 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5401 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5461 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5521 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5581 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5641 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5701 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5761 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5821 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     5881 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     5941 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     6001 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     6061 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     6121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     6181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     6241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
     6301 cagcagccat catca
//
