LOCUS       sequence-317767-        6291 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 6291)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6291)
  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_017947
FEATURES             Location/Qualifiers
     source          1..6291
                     /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             1112..1129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1182..1200)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1196..1243
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1280..1735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1367..1386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1577..1598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1827..2642)
                     /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(1827..2642)
                     /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     2550..2569
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2634..2653
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2764..3352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3253..3272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3506..3523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3538..3677
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3603..3622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3763..3785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3779..3970)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4779..5861)
                     /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     5823..5842
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5862..5939)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6145..6164
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6248..6267
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6248..6266
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6267..6291
                     /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 catatgcaac tgcgtcgtgt
      121 acgaatagca ggcacgggta aatatctgcc gcagatgaag gtaaccgatg aagaactgga
      181 cgaaaggtta catgtcccgg cgggctgggt gtctaaagct acgggcgttg gttaccggta
      241 ctacgcgtct gatgaagaga cctcaagcta tatgggagca cgggccgccg aagcggcgtt
      301 ggcggatgca ggactgagtt ttggtgagat cgattgcttg gtgtgcactt ccggaacaaa
      361 ggaacaacct ttaccgtcaa cggccgtttt tatacagcag gctatggggc aggaggagtc
      421 gggtgtacct gcctttgata tggatgcgac gtgtttatcg tttttaaatg gtctggacgt
      481 gatctcctat atggtcgacg cgggccgcta tcagcgtgtg ttgctcgttg caactgagat
      541 tgccagtgcc ggcctgaact ggaaagatcc tgaatccgcg gccctctttg gcgatggtgc
      601 agctgcagtc attattgaac gagcgtctga ggaatctggc tcacagatag ttcatgcttc
      661 tctgcgtacc tactcgcgtg gcgctcgctt cagcgaaatc gcaggcggtg gtactaggca
      721 tcacgcggct caatatgatg ttgcccatcc tgatccattt ctttttcaca tggatggaca
      781 ggccatattt aggatggctt ccaaacttct tcctggattt atgaatgaaa tgctccaggc
      841 ttctggtgac cgtatggaag acttttcact ggtgatcccc caccagggct cggcgatggc
      901 aatgcgatta ataagaaaaa agctagggat acccgctgaa cgctttatgg atattaccca
      961 gggccacggg aatacgatag ccgcgagcat tccgatgggc cttcatgaag ccattcgcca
     1021 gcagcgtatt gaaagaggtg accgtgtttt aatgattgga acggcggcgg gcctgtcttt
     1081 gggtggctta atcttcgatt actaactcga gcaccaccac caccaccact gagatccggc
     1141 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1201 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1261 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1321 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1381 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1441 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1501 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1561 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1621 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1681 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     1741 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1801 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     1861 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1921 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1981 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     2041 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     2101 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2161 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     2221 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2281 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     2341 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2401 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2461 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     2521 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2581 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     2641 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     2701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2881 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3541 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     3601 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3661 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3721 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3781 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3841 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3901 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3961 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     4021 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     4081 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     4141 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     4201 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     4261 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     4321 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     4381 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     4441 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4501 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4561 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4621 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4681 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4741 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4921 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4981 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     5041 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5101 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5161 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5221 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5281 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5341 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5401 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5461 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5521 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5581 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5641 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5701 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5761 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5821 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5881 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5941 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     6001 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     6061 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     6121 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     6181 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     6241 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc c
//
