LOCUS       sequence-317182-        6303 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 6303)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6303)
  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_018063
FEATURES             Location/Qualifiers
     source          1..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             719..736
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(789..807)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      803..850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      887..1342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(974..993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1184..1205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1434..2249)
                     /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(1434..2249)
                     /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     2157..2176
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2241..2260
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2371..2959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2860..2879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3113..3130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3145..3284
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3210..3229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3370..3392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3386..3577)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4386..5468)
                     /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     5430..5449
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5469..5546)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5752..5771
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5855..5874
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5855..5873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5874..5898
                     /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             5913..5935
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5954..5971
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5981..5998
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 ctcaattgta catcgtcatc taggactgag cccgcgatgt cgtaattttg atttagcgaa
       61 tgcgtgccta ggtttcctgg acgggatgga acttgtggcc gcgatgattg aacgcggcgc
      121 gatagatgcc gccctggttg tcgacggtga gggctccaga tttgctgttg aaaaaacaat
      181 tgatagactg aacgcgagcg gtgatgcacg tgcactgggt gaagaattcg cgactctgac
      241 actgggttcc ggggctgctg ccatggttct gtgtcgccga gacttggctc cagctgctca
      301 tgcttttaga ggaggcgtat ctttagctgc gaccgattgg aatcatttat gtcggggtca
      361 tgcagatcgc atggttaccg atgcccagaa gctcctgacc caaggactgg accttgcttc
      421 aaaaacgttt gaattagcgc gtactgaact cggatggtcg cctgatgttc tcgacgaggt
      481 ggtgattcat caggtgtctc gacaacatac agaaaaattg gctgcacagc tgggtgttgc
      541 acccgctaag attcatgcaa tttttcctga acatggcaac gttggtccgg caagtgtgcc
      601 tatggtatta gcgaaagccg cagaaattgg cagattacgg agcggcatgc gtgttgccct
      661 tatgggcatt gggtcgggtc ttaattgcgc tatggccgaa attgtttggt gactcgagca
      721 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
      781 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
      841 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
      901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1021 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1321 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1381 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     1441 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1501 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1561 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1621 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1681 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     1741 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1801 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1861 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1921 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     1981 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2041 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2101 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2161 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     2221 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2281 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3121 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3181 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3241 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3301 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3361 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3421 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3481 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3541 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3601 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     3661 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     3721 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     3781 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     3841 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     3901 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     3961 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4021 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4081 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4141 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     4201 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     4261 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4321 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4501 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4561 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4621 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     4681 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4741 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4801 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4861 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4921 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4981 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5041 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5101 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5161 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5221 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5281 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5341 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5401 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5461 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5521 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5581 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     5641 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     5701 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     5761 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5821 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     5881 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     5941 catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc gcggcagcca
     6001 tatgcttttt tcctccgtga gcgtagcctc agtggcctat gttgatgcgc ctcatcgggt
     6061 gtccagtcgg gagctggaag ataggttatc tccgcttctt aaaaggctgg gcctggcccc
     6121 aggcataatc gcttccttat ccggagtgca agctcgcagg ttttgggatg agggagttca
     6181 accatctgaa gtagccgcac aagcaggtga gaaagcgatt gtagcgtccg gtatcgatcg
     6241 cgcgcgtatt ggggtgcttg tgaacacctc tgtatgccgt gacttcgtag agccatctac
     6301 agc
//
