LOCUS       sequence-315979-        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_018101
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     286..303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    318..457
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(383..402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     543..565
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(559..750)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1559..2641)
                     /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     2603..2622
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2642..2719)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2925..2944
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3028..3047
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3028..3046
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3047..3071
                     /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             3086..3108
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3127..3144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3154..3171
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4207..4224
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4277..4295)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4291..4338
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4375..4830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4462..4481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4672..4693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4922..5737)
                     /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(4922..5737)
                     /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     5645..5664
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5729..5748
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5859..6315,1..132)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
       61 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      301 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      361 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
      421 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
      481 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      541 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
      601 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
      661 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
      721 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
      781 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
      841 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
      901 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
      961 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     1021 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     1081 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     1141 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     1201 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     1261 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1321 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1381 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1441 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1501 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     1561 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1621 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     1681 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     1741 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     1801 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     1861 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     1921 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     1981 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     2041 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     2101 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     2161 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     2221 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     2281 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     2341 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     2401 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     2461 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     2521 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     2581 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     2641 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     2701 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     2761 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     2821 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     2881 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     2941 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     3001 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     3061 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc
     3121 agcagccatc atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatgtct
     3181 ggcaatgcta gttatcacct gagtaactgt tccatcttgg cggtgactac agtggaggcg
     3241 ccagaaattg tcactagcgc agaattagat accgcgttgg ctccagctta taccgaaacg
     3301 cgtaccgtgg ccgggcagtt agaacgtctg gcaggtgtaa agcaacgccg ttggtttccg
     3361 gccgacgaag atttcaccga tggcgcggtt gaggctggac gtcgcgcact acacacagct
     3421 ggcatatccc cagagcaggt gggtttagtg ataaatgctt ccgttacccg ccctcatctg
     3481 gaaccgggta tttcggctaa agtccatgct gaacttggcc tgccccgaaa ctgcatggat
     3541 ttcgacgtga ccaacgcatg cctgggagta gtaaatggaa tgcagatcgc aggtacaatg
     3601 atagacgcgg ggcaaattcg ttatggcctg attgttgctt cggaatccag ccggcaaatg
     3661 caagaagcaa caattaagcg gcttaatgcg ggaggtgcaa ctaaatccga tgtaaagcaa
     3721 gcgtttgcca ccatgacgct gggctcgggt gctgttgcca tacttatcgg tccgacagat
     3781 gctcaccctg aaggacatcg gttattaggt ggtatcaccc gagccggaac ggagcaccac
     3841 agactgtgcg tgggttccat ggatgggatg agaacggaag catcaaaact gtttcgagag
     3901 ggaattgact tagctctgga cgcgtatacc gagagtgctg aagatgggtg ggactggaag
     3961 gacatggatt ggtatgtggc acaccagacc tcaacagcgc atattgcgac ccttgctcag
     4021 cgtctgggaa aaccgctgga aaaatttcca accacaatta gtgaatatgg aaatattgga
     4081 ccagctgcaa tgccgtatac tctgggtaca ttccaggatc aatttgcaac cggagatcgt
     4141 ctgcttctca tgggcatagg gtccggtttg aatgcctctt tcgtggaggt tcagtggtaa
     4201 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     4261 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4321 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     4381 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4441 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4501 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4561 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4621 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4681 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4741 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4801 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     4921 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     4981 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     5041 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     5101 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     5161 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     5221 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     5281 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     5341 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     5401 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     5461 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     5521 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     5581 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     5641 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     5701 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     5761 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6001 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6301 gtaagcggca gggtc
//
