LOCUS       sequence-315999-        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_018117
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             519..536
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(589..607)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      603..650
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      687..1142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(774..793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     984..1005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1234..2049)
                     /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(1234..2049)
                     /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     1957..1976
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2041..2060
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2171..2759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2660..2679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2913..2930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2945..3084
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3010..3029)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3170..3192
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3186..3377)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4186..5268)
                     /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     5230..5249
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5269..5346)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5552..5571
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5655..5674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5655..5673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5674..5698
                     /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             5713..5735
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5754..5771
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5781..5798
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 ctccaggggt cactcgcgcc gacgttgtaa gggaatttgc aacactgacc ttaggctgcg
       61 gtgcagcagc tgcggtggtt ggtccagcag atcgtcaccc cgaaggacat agagtggcgg
      121 ggagtgttac gcggtccgcc acagagcatc atggtctgtg tattggtgga tttgatggca
      181 tgtttacaga tgcaacaggt ctgctgaagc atggtgtggg gttactcgaa caggcttgga
      241 acgatgctca tgcagatgga tgggattggc aggagatgga tcgctacatc ggacaccagg
      301 tgtccatgtc acatacgaat aaactgttcg ctgacgtcgg aattgatccg gacaggtttc
      361 cagttacatt ccctttttgg ggaaatgtgg ctgcagcggc gttgccaatg accttagctc
      421 tagaagccga aagtctgggt gcgggtgaca gagtgctcct tttgggtgtt ggatcgggtc
      481 ttaatgcaac ttatatggaa atcgactggt gactcgagca ccaccaccac caccactgag
      541 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      601 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      661 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
      721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
      781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      841 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
      961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1021 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1081 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1141 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1201 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     1261 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     1321 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1381 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1441 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1501 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1561 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     1621 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     1681 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1741 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1801 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1861 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1921 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1981 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2041 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2101 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2161 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2221 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2281 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2341 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2401 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2461 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2521 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2581 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2641 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2701 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2761 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2821 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2881 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2941 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3001 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3061 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3121 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3181 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3241 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     3301 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     3361 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     3421 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     3481 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     3541 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     3601 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     3661 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     3721 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     3781 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     3841 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3901 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3961 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4021 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     4081 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4141 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4201 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4261 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4321 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4381 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4441 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4501 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4561 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4621 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4681 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4741 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4801 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     4861 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4921 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     4981 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5041 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5101 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5161 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5221 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5281 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5341 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5401 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5461 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5521 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5581 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     5641 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     5701 tctagaaata attttgttta actttaagaa ggagatatac catgggcagc agccatcatc
     5761 atcatcatca cagcagcggc ctggtgccgc gcggcagcca tatgacgggt aattcaacaa
     5821 tgagacatcg gaacgcggca ctgttagcga tcactaaagt tgaagctgat gaagttgttc
     5881 cgtcttcgca agcagatgaa gttttggccg gcacgtttga acgcttacgt atgcctaaag
     5941 gcctgctgga acgactggcc ggcgttaaag aacgtcgtga atgggcccca ggcaggcatt
     6001 tcactgacgg cgctgtagag gcaggtcagg cggcgcttga gcaagccggt gtgcgtccag
     6061 agcaggttgg attactgatt aacgcctccg tcacacgcga tggctatgaa cctgcagtca
     6121 gcgtaagcgt gcatcatcgt ttaggcctgc caagctcggc actcaatttt gacgtcacaa
     6181 acgcctgttt agggtttgtc aatggcctga ctattgctgc taccatgatt gatgcaggtg
     6241 ccatcgatta tgccgtggtc gtcgcaggtg aagacccccg accatggcat agagagacgt
     6301 atgagcgcct gcagg
//
