LOCUS       sequence-317450-        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_017967
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     973..992
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1012..1089)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1295..1314
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1398..1417
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1398..1416
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1417..1441
                     /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             1456..1478
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1497..1514
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1524..1541
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2577..2594
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2647..2665)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2661..2708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2745..3200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2832..2851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3042..3063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3292..4107)
                     /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(3292..4107)
                     /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     4015..4034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4099..4118
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4229..4817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4718..4737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4971..4988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5003..5142
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5068..5087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5228..5250
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5244..5435)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6244..6315,1..1011))
                     /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"
ORIGIN
        1 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
       61 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      121 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      181 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
      241 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      301 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      361 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      421 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      481 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      541 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      601 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      661 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      721 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      781 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      841 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
      901 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      961 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1021 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1081 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1141 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1201 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1261 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1321 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1381 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     1441 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc
     1501 atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatgcgt gggaacgcta
     1561 ctacacgcca ctctaatgta gcgctgcttg gtataacgga ggcggtcgct cccgttgagg
     1621 tgagcagcaa gagttttgat gatcgtttag ccgacacact gaaaaccttg cgtttaccca
     1681 aaggattgct gcagcgtgtc gccggtgtta ccacacgccg caattgggaa cacccttccg
     1741 attacatcga gggcgccgca gatgctggcc gcagagcgct ggaagaagct ggcgttcgcc
     1801 cggatcagat tggtctgctg attaacgcga gtgtgagccg tgaaatgttg gagccgagcg
     1861 tcgcggtggc ggtgcatgac tctattggtc tgagtcccgc cgcgatgaat tttgatatta
     1921 ctaatgcatg cctgggtttt gtgaatggta tgacggtagc agctagtatg atcgatgcag
     1981 gccagatcga ctatgctctg attatcgccg gggaagacat gtctcgcgtc caggagtcta
     2041 ccgtggaacg actacgaaga gacggtatta cgcgtgacga atacttgaat gaattcgcga
     2101 cgttaacctt gggaagcgga gcagcggcag ctgtgcttgg gcgtgcggat cgtcacccgg
     2161 aaggtcatct gattaaaggt ggagttacac gcgccgcgac atggaatcat ggcctgtgta
     2221 ttggcgatta cgacggtatg ttcactgaca ctcaaggtct tcttgacaac ggcatgcagc
     2281 tgatctctga agcatgggca gaggcccacg ataacggatg ggaatggacc acaatggacc
     2341 attatatacc ccaccagatc agccatgtgc atgtaaatgc gctaaccgag gctacgaaaa
     2401 ttgatgcaga tcgcgtcgcc aaaacatatc ctgagctggg taatgtagga ccggcgagcc
     2461 tgccgataac actggcacga aaatccggtg agatgaagca gggtgatcgg atcctgtgtc
     2521 ttggcgtggg gagtgggtta aataccgcta tgttggaaat cacgtggtag ctcgagcacc
     2581 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     2641 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2701 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     2761 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2821 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2881 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2941 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3001 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3061 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3121 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3181 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     3301 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3361 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3421 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3481 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3541 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3601 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3661 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3721 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3781 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3841 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3901 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3961 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     4021 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     4081 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     4141 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4981 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5041 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5101 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     5161 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5221 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     5281 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     5341 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     5401 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5461 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5521 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5581 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5641 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5701 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5761 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5821 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5881 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5941 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     6001 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     6061 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     6121 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     6181 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     6241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6301 caacgcgcgg ggaga
//
