LOCUS       sequence-316879-        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_018105
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    127..266
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(192..211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     352..374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(368..559)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1368..2450)
                     /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     2412..2431
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2451..2528)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2734..2753
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2837..2856
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2837..2855
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2856..2880
                     /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             2895..2917
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2936..2953
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2963..2980
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4016..4033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4086..4104)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4100..4147
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4184..4639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4271..4290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4481..4502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4731..5546)
                     /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(4731..5546)
                     /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     5454..5473
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5538..5557
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
      181 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      241 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
      301 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      361 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
      421 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
      481 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
      541 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
      601 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
      661 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
      721 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
      781 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
      841 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
      901 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
      961 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     1021 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1081 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1141 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1201 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     1261 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     1321 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     1381 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1441 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     1501 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     1561 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     1621 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     1681 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     1741 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     1801 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     1861 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     1921 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     1981 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     2041 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     2101 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     2161 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     2221 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     2281 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     2341 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     2401 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     2461 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     2521 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     2581 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     2641 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     2701 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     2761 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     2821 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     2881 cctctagaaa taattttgtt taactttaag aaggagatat accatgggca gcagccatca
     2941 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatgtctg gcaatgctac
     3001 ttataggctg tctaatacga ctatcctggc cgcctgcgca gtagaagccc ccatcgtcgc
     3061 cacctccgca gccttcgatg agcgattggc tgaaacatac cgccgcgtgg gcacccgccc
     3121 tggtctgctg gtaagactgg caggtgttca ggaacgtcgt tggtggccgg aagacgtaac
     3181 ttttgccgat gcagccgcaa tggcaggcgc taaagcattg gctgaggctg ggattcgtcc
     3241 ggagcaggtc ggcgtggtga ttaatacttc cgtctgcagg tcccacctcg aacctagcgt
     3301 tgctgtggcg gttcatcatc agctggggct cccgactaac tgcttgaatt tcgacgtggc
     3361 caatgcgtgt ttaggttttg tgaatgcgat tcaattagct ggaacgttga tagattctgg
     3421 gcagttggac tacgcgctga tcgtagatgc tgaggggagc agacaaattc atgagctgac
     3481 cttagatcgc ctgtcgagtc ctacggcgac ggcggcagaa tttaaggaac agttcgccag
     3541 cctaacgatt ggttccggtt ctgccgcctt tgtgctcggg agagcagatc gccatccaga
     3601 aggtcatcgc gtcattgggg gggctagccg aaccggttct gaacaccacg atctgtgtgt
     3661 tggtagtctc gatatgatgc gcacggattc taagaaactg tttgaccacg gcattgcact
     3721 tgcagtcgct acatgggaag atgcaaaaga tgagttcgat tgggacggtg ccgttgattg
     3781 gtttattgcc catcagacaa gtacagtaca cttgcgcgct atggcgaaag ccgtaggcgt
     3841 cagcagtgca cactttccga ccacagttga aaaatatggc aatatgggtc cagcagccgt
     3901 gccgttcaca ctggctcatc atgccgatcg catgcgagca ggcgatcgca ttgttctgat
     3961 cggcatcggg agcggactga acacgagctt cgcagagatt gtttggtgac tcgagcacca
     4021 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     4081 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     4141 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     4201 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4261 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4321 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4381 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4441 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4501 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4561 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4621 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     4741 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     4801 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     4861 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     4921 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     4981 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     5041 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     5101 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     5161 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     5221 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     5281 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     5341 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     5401 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     5461 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     5521 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     5581 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6301 gccttttgct cacat
//
