LOCUS       sequence-316000-        6318 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 6318)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6318)
  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_018127
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    76..100
                     /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             115..137
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             156..173
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             183..200
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1239..1256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1309..1327)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1323..1370
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1407..1862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1494..1513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1704..1725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1954..2769)
                     /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(1954..2769)
                     /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     2677..2696
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2761..2780
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2891..3479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3380..3399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3633..3650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3665..3804
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3730..3749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3890..3912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3906..4097)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4906..5988)
                     /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     5950..5969
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5989..6066)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6272..6291
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
       61 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
      121 taactttaag aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg
      181 gcctggtgcc gcgcggcagc catatgatac cgaacaccgc ctttgatcac aaaaatgccg
      241 ccattttgtc ggtgtcagcc gtcgaagcgc aggtgccggt tacctcagat tcaatcgatg
      301 aaatgctgag tccggccaga aaacgcctcc gcttgccaaa ggggaccttg cagcgtgttg
      361 caggtgtata cgagcggcgg tggtggaaag atccggagaa tggatggaag cagggagcag
      421 tgattgctgc cgaacgcgcc atggcgtcgg ctggtatcac acgggaggat gtaggcgtct
      481 taataaacgc ttccgtttcg agacaacatc tggaaccggc catgagtgcg catatacacg
      541 ctgccctggg catgccggct agttgtatga attttgatat tacaaacgct tgtctcggat
      601 ttgttaacgc catgaccctt gcagctgggt tgattgacgc gggtcagatt gattatgcat
      661 tgattgtcgg ggcagaagat gttgaagagg tgcaacgcag gactatagca agactgaacg
      721 acaaaaattc aactaggcaa gacttcaata atcagtttgc aagtctgaca ttagggagcg
      781 gtgccgctgc ggcggtagtt ggccggtttg acaagcatcc ggatggacat agaattgttc
      841 acacacagac gcgggcgggg acagagcatc atgagttatg cattggaaat atggatgata
      901 tgagaacaga taccgctggc ctgctggaaa acggactcga acttgtgaca cagacatgga
      961 gagcagccga ggaaagtggc cgtaactatt ctaataaaca atggtacatc acacaccagg
     1021 tgagcatggt atatacgcgt gcatttgcca aagccatggg gatcaacccc gaaattatcc
     1081 ccgttacctt tccggaacta ggaaatatcg cagctgaagc cgttccgttg actctggcat
     1141 tgaaccagga taggtttgca aaaggtgacg agctgttact cttaggagta ggatctgggc
     1201 tgaatacagc tatgatggaa gtgcagtggt gactcgagca ccaccaccac caccactgag
     1261 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1321 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1381 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1441 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1501 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1561 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1621 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1681 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1741 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1801 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1861 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     1981 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2041 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2101 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2161 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2221 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2281 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2341 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2401 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2461 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2521 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2581 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2641 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2701 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2761 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2821 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3181 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3241 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3301 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3361 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3421 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3481 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3541 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3601 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3661 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3721 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3781 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3841 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3901 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3961 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4021 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4081 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4141 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4201 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4261 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4321 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4381 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4441 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4501 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4561 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4621 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4681 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4741 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     4801 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4861 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4921 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4981 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5041 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5101 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5161 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     5221 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5281 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5341 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5401 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5461 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5521 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5581 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5641 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5701 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5761 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5821 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5881 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5941 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6001 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6061 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6121 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6181 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6241 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6301 ccgcacctgt ggcgccgg
//
