LOCUS       sequence-316868-        6327 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 6327)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6327)
  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_018139
FEATURES             Location/Qualifiers
     source          1..6327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             81..98
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             108..125
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1173..1190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1243..1261)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1257..1304
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1341..1796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1428..1447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1638..1659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1888..2703)
                     /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(1888..2703)
                     /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     2611..2630
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2695..2714
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2825..3413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3314..3333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3567..3584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3599..3738
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3664..3683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3824..3846
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3840..4031)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4840..5922)
                     /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     5884..5903
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5923..6000)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6206..6225
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6309..6327,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6309..6327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
      121 gcagccatat gcggtatcag cgtgttttta ttaacaaaat cgcttatgag ctcccgaaag
      181 aaaaggtggc gacttccttt ctggaagagc agttaaccga cgtgtatcaa gagctcggta
      241 tcccactagg ccaggtcgaa gctctgacaa aaatccatca gcgtaggtac tggcctgtgg
      301 accatcaact gtctgatcat gcagcgctgg cagcaaaaaa agccattaaa gaatcgggaa
      361 ttaatagtaa aaaaatccag gtgttaattt acgcaggagt aggtcgagca cagctcgaac
      421 cggcgacggc ttgtgccgtt gcacaccaac tcgcaattgc ttcggatgca tcagtttacg
      481 atgttagcaa tgcctgttta ggcgtgttaa acggtattat tcaaattgcc aatgcaatag
      541 agctaggtca gatccatgct ggtctagtgg tctctgcgga gtcgtgtcgc gagattatag
      601 atacaatggt cgcggaaatg aaaattcagc gaaacataga gcattttgca ctgagccttg
      661 ctacactgac gggagggagc ggcgctgttg ccgttctgct ttcagataat gcgttagatt
      721 caagtcacca gctgctgggc gctgtagcaa ggcagaaatc acagtggcat caattgtgcc
      781 gttggggctc taagacgggt attttgggaa atgcatgtaa tatcatgcat acagattccg
      841 tttcagtact gaagtacggt gtggaactgg gtgtagaaac ctataaggat tttttgcaag
      901 aattgttatg gcaggacagc ttccctgata aatatatatg ccatcaagtc ggtgaggcaa
      961 atcagctcgc tattcgtcgc gcattaaaca ttccgaaaga aagagatttt acaacgtatg
     1021 aaacactggc taacactggt acagtggctc tgccgatgac tgctgcaatt gcagatcagg
     1081 taggtttcct gaagaccggc gataaggttg cgtttctggg tataggttcc ggcctcaatt
     1141 gcatgatgct tggtattaaa tggtaactcg agcaccacca ccaccaccac tgagatccgg
     1201 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     1261 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     1321 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     1801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1861 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     1921 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1981 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     2041 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     2101 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     2161 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2221 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     2281 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     2341 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     2401 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     2461 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     2521 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     2581 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2641 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     2701 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2941 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3481 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3541 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3601 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3661 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3721 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3841 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3901 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3961 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4021 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4081 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4141 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4201 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4261 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4321 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4381 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     4441 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     4501 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4561 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4621 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4681 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     4741 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     4801 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4861 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4921 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     4981 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5041 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5101 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5161 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5221 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5281 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5341 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5401 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5461 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5521 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5581 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5641 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     5701 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     5761 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     5821 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     5881 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     5941 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6001 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6061 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     6121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6181 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6241 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     6301 cgcgaaatta atacgactca ctatagg
//
