LOCUS       sequence-316878-        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_018109
FEATURES             Location/Qualifiers
     source          1..6327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(53..130)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     336..355
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     439..458
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        439..457
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    458..482
                     /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             497..519
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             538..555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             565..582
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1630..1647
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1700..1718)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1714..1761
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1798..2253
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1885..1904)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2095..2116
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2345..3160)
                     /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(2345..3160)
                     /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     3068..3087
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3152..3171
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3282..3870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3771..3790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4024..4041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4056..4195
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4121..4140)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4281..4303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4297..4488)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5297..6327,1..52))
                     /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 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
       61 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
      121 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
      181 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
      241 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
      301 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
      361 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
      421 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      481 cccctctaga aataattttg tttaacttta agaaggagat ataccatggg cagcagccat
      541 catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatgca aaaacggagt
      601 gagggcaata ctactttcca ccatcggaac gcggcactgt tgagcgtcgc cgaagttgaa
      661 gcccccttag ttttgacgag taaagacttg gataaacgac tggctgacgc cctgaaaagg
      721 ctaaaattac cgtcagggct gttgcagcgg gtagccggcg tgaaagccag acggaattgg
      781 cgttctgatg acgaagtgcg cgccggaact gttgaagcag gacgtcgcgc gttggaagcg
      841 gccggcgtct caacagaaga catctcagtg atgattaata cttcagtgac gcgcgaacat
      901 cttgaaccgt ctgtggccgt tggtgtgcat catgagctgg gcctgccatc cagtgctatg
      961 aatttcgata ttacgaatgc ctgcctggga tttgtcaacg cgatgactct tgccgcaact
     1021 atgattgatg caggtcaggc ccgatatgta cttattgtta acggagaaga tgcacgtaaa
     1081 gttcaagatg ccacggtcga acggataaat gcccttgagg aagtttcacg tcaggatttt
     1141 atgactgaat tcgcgtcgct gacccttggc tgtggagctg ccgctgccgt aatcggtccg
     1201 gccgacgagc acccagaagg tcaccgtatc gtcggcgggg tcactagggc ggcgacgcag
     1261 caccacgaac tatgtgtagg cgatcaccgt gggatgttca cggacgcgcg cggcctttta
     1321 gatggtggac ttgaattagt gatgaacgcg tgggaagacg ccaaaaaaca tttttcctgg
     1381 caaagtatgg acgcctacgt gactcatcag gtgagtcaac tacatacgag ttccattatt
     1441 aaggcagcca aactggataa gaaaaaggtt ccagttacat atcctgaact tggtaacgtt
     1501 ggtcccgctt ccttacccat aacacttgca cgtgaggcca agaatctggt tactggttcc
     1561 cgtgtcctgt gtatgggcgt tggttcaggt ctaaacaccg ctatggtgga gattgagtgg
     1621 tgactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     1681 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1741 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     1801 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1861 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1921 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1981 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2041 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2101 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2161 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2221 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     2281 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     2341 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     2401 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     2461 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     2521 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2581 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     2641 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2701 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2761 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2821 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     2881 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2941 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3001 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3061 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3121 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3181 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     3241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3421 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4081 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     4141 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     4201 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4261 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4321 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4381 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4441 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     4501 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     4561 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4621 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     4681 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     4741 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     4801 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     4861 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     4921 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     4981 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     5041 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     5101 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     5161 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     5221 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5281 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     5401 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5461 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5521 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     5581 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5641 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5701 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5761 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5821 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5881 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5941 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     6001 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     6061 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6121 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6181 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     6241 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     6301 cctggttcac cacgcgggaa acggtct
//
