LOCUS       sequence-317629-        6300 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 6300)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6300)
  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_017953
FEATURES             Location/Qualifiers
     source          1..6300
                     /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             1146..1163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1216..1234)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1230..1277
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1314..1769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1401..1420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1611..1632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1861..2676)
                     /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(1861..2676)
                     /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     2584..2603
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2668..2687
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2798..3386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3287..3306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3540..3557
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3572..3711
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3637..3656)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3797..3819
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3813..4004)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4813..5895)
                     /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     5857..5876
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5896..5973)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6179..6198
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6282..6300,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6282..6300
                     /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 gtcagcgccg cgttactcac aggtatctgc cgttgccgta cgtctaccag
      181 acgaagattt aactaccccc gaattggagg aattgttagc agaacgcaat ccccgagtcg
      241 atgttccgcg cggacttatc gaaagactct caggagtccg ccgccgacac gtcgcaccac
      301 cgggtgtgct gccctccgat ttagcagtgg cggctgcacg tgacgtactg cgccaaaccg
      361 gccatgcacc gcccgcaatc gacctgatta tttatgccgg tgtgtcggct gacgccgtag
      421 aaccggcaac cgggcacatg gttgcagcga agcttggggc cgactgtcct gtttttgatg
      481 tgcgcaatgc ctgcaacagc gttctgaacg caattgaggt tgcagacctc tttattactt
      541 caggccgaca tcggactgtg ctgattgcct gtggcgaggt cattactgca gcgatgcctt
      601 ggcgactcgc ctctgaggaa gagtttacga ccgcggtgcc aagctacact gtctctgatt
      661 ctgggtcagc actcctgctg caggctgctg acgccccagg cgtacttggc catcgcttct
      721 gcgcaaacag cgctgattgg gaagttagcg cgatgccttt cgttagagat ggcgccggag
      781 ccctgcgggt tggcgcattt actattgatt ccgttggttt ggcgatcggg gttcaaaaac
      841 tggataggga aatttggaga aggccacttg cagaacatgg tctggactgg gatgatatgg
      901 cagcaatatg cgtgcaccaa ccgagcttag cagctctggg ggcggtgtgt gagttagcgg
      961 gaatccctcg tgataaagcc gaagtaacca ttgcagatca cggtaactta gctgcatcta
     1021 caataccgat gcaactgcac ttggcaacag aggccggtcg cgttcgtcgg ggcgacctgg
     1081 ttgctcttgt gggagttgcc agtgggattt ccgctgggac cgtcctgctt cgctggtaac
     1141 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg
     1201 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1261 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     1321 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1381 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1441 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1501 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1561 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1621 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1681 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1741 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1801 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     1861 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     1921 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     1981 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     2041 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     2101 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     2161 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     2221 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     2281 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2341 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     2401 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2461 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     2521 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     2581 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     2641 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     2701 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     2761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2941 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3421 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3481 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3541 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3601 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3661 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3721 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3781 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3841 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     3901 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     3961 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4021 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4081 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4141 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4201 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4261 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4321 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4381 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4441 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     4501 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4561 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4621 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4681 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4741 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     4921 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     4981 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5041 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5101 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5161 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5221 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5281 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5341 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5401 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     5461 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5521 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5581 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     5641 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     5701 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     5761 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     5821 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     5881 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     5941 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6001 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6061 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6121 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6181 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6241 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
//
