LOCUS       sequence-316877-        6318 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 6318)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6318)
  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_018111
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    124..263
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(189..208)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     349..371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(365..556)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1365..2447)
                     /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     2409..2428
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2448..2525)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2731..2750
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2834..2853
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2834..2852
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2853..2877
                     /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             2892..2914
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2933..2950
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2960..2977
                     /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 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
      181 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
      241 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
      301 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
      361 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
      421 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
      481 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
      541 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
      601 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
      661 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
      721 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      781 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
      841 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
      901 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
      961 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     1021 gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     1081 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     1141 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     1201 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     1261 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     1321 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     1381 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1441 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     1501 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     1561 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     1621 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     1681 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     1741 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     1801 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     1861 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     1921 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     1981 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     2041 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     2101 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     2161 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     2221 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     2281 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     2341 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     2401 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     2461 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     2521 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     2581 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     2641 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     2701 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     2761 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     2821 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     2881 ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca
     2941 tcatcatcac agcagcggcc tggtgccgcg cggcagccat atgtcagaag cgatacgcgc
     3001 acgcattgcc gcagtatgcg cccatcttcc aagtcggtat cgtaccctgg atgagactcg
     3061 ggcaaagatt acagagacgg gaggttatgc tccgccgccc ggactgttag aggatcttac
     3121 aggggtgcgt ggagtccatg ttcatgaaga tggtgagaat tcaagtgatc tggcggtggc
     3181 agccgctagg agcgcactgg agcaggcagg acttaaaatt cgagatgttg acttactgtt
     3241 attcgcgtcg acttcacagg atttgatcga gccggctacc gcccaccttg ttgcaaccaa
     3301 acttggagcc agctgtccag tgtttgacgt caaaaatgca tgtaacagct ttctgaatgg
     3361 tatggaagtg gcagcgtcct tcattgagac ggggcggtat cggacggtcc ttattgcttg
     3421 tggggaaccc gctaccctag cgactagatg gcatttgccc acacgcgaag atttactgag
     3481 agcattgcct ggatataccg cttctgatgc cggtgctgca atggtgctca cagctggccc
     3541 ggcgggtgat ggcgaccccg gaattcttag cctccgcttt gtctcaaaca gtccggcctg
     3601 ggaagcgtgt acggttgcag ggggtggctc tatgcatcat ttgagcgttc atgatgacca
     3661 tacgaccctg cgactaaatg gtgacctctg gcagtcagcg ctcgatgtgt taccgagaat
     3721 catggataca gcaggtgagg agtttcgtgc agcccaggct ggcgcagtgt ccttcatcgc
     3781 ctttcaccag attgcaatgc cgcaatatcg cgaagttatt gatcttttct ccttacctga
     3841 ggatcgctgt atgccagcag tggcggagca cggtaactgc gcggccgcgt ctctgccgct
     3901 gcaattagcc aaagcccgcg aggcggatcg tattgagggg ggtgatctgg tcgccatgat
     3961 aggcttggca tcagggttct cattcggact tgccttggta cgctggtgac 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 cacatgtt
//
