LOCUS       sequence-317634-        6279 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 6279)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6279)
  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_017937
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     948..967
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(987..1064)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1270..1289
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1373..1392
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1373..1391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1392..1416
                     /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             1431..1453
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1472..1489
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1499..1516
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2513..2533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(2586..2604)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2600..2647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2684..3139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2771..2790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2981..3002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3231..4046)
                     /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(3231..4046)
                     /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     3954..3973
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4038..4057
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4168..4756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4657..4676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4910..4927
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4942..5081
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5007..5026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5167..5189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5183..5374)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6183..6279,1..986))
                     /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 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
       61 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
      121 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
      181 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
      241 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
      301 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
      361 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
      421 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
      481 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
      541 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
      601 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
      661 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
      721 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
      781 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
      841 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
      901 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
      961 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     1021 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     1081 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     1141 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     1201 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     1261 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     1321 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     1381 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     1441 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
     1501 ggtgccgcgc ggcagccata tgggtactca gctggcagga gttgccgtac acttacccga
     1561 gcaacaggtg tcgacgaccg aggtggagcg tcgtctagct ccttttactc caccagccgg
     1621 cctgatccac cgcttaacag gtgtggaatc tcgacacgtg atgggtgaac atgaacaagc
     1681 atcagatctg gctgttgcgg ctgcccgaaa attgctggcc gaaacgggaa cggcagcagc
     1741 ggatgtggac ttgcttcttt ttgcaagcgc gtctcaggat atggttgaac cagccaccgc
     1801 ccatattgtt gctgccaaac tggaactgcg ttgtcctgta atggatgtca aaaatgcctg
     1861 taatagtgtt cttaatggta ttgaagtagc ggatgcttta attgcatctg gtagatatcg
     1921 taccgtgctc gttgccagcg gcgaagcccc gagcagagct gtacgttggc aggtgggcgg
     1981 gctggctgaa tttttgagag cctttccggg gtacacctta agtgacgccg gcgcggcact
     2041 gctgctgact cgcggtgaag aaggggttct gggctgtggt ttcacagcgg acagctcacg
     2101 ttgggacgtg ggcacgcttc caggtggagg ctctcgttat ccccgcgatc cagagcgaac
     2161 atattttgat atggatggcg cccgcctgaa ggatgcattt ctcggccttg gtcgtggcgt
     2221 tttagatagc actctggacc ggcttggcct gacttgggat gacttcgccg taattggtgt
     2281 acaccaggtt tctcttccgt atcttgatat gtttgcgcgc caggtcggtg ttcctactga
     2341 tcggctggtg attactttac cacaacacgg taacgttgcg agcgccagtt tgcctttaca
     2401 attggtgacg gcgcggcagt tgggacgctg cggtccaggc gatctggtag cgctggtggg
     2461 attagctgga ggtgtcagcc tgggcttagt cgtgattcgt ttgtgactcg agcaccacca
     2521 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2581 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2641 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     2701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3121 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     3241 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     3301 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     3361 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     3421 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     3481 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     3541 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     3601 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     3661 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     3721 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     3781 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3841 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3901 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3961 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     4021 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     4081 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4741 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4801 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4861 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4921 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4981 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5041 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     5101 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5161 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     5221 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     5281 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     5341 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     5401 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     5461 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     5521 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     5581 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     5641 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     5701 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     5761 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     5821 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     5881 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     5941 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     6001 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     6061 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     6121 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     6181 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6241 aacgcgcggg gagaggcggt ttgcgtattg ggcgccagg
//
