LOCUS       sequence-316778-        6363 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 6363)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6363)
  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_017991
FEATURES             Location/Qualifiers
     source          1..6363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     958..977
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(997..1074)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1280..1299
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1383..1402
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1383..1401
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1402..1426
                     /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             1441..1463
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1482..1499
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1509..1526
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2610..2627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2680..2698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2694..2741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2778..3233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2865..2884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3075..3096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3325..4140)
                     /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(3325..4140)
                     /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     4048..4067
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4132..4151
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4262..4850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4751..4770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5004..5021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5036..5175
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5101..5120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5261..5283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5277..5468)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6277..6363,1..996))
                     /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 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
       61 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
      121 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
      181 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
      241 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
      301 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
      361 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
      421 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
      481 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
      541 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
      601 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
      661 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
      721 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
      781 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
      841 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
      901 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
      961 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     1021 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     1081 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     1141 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     1201 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     1261 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     1321 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     1381 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
     1441 tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca
     1501 gcagcggcct ggtgccgcgc ggcagccata tgaaattctc acagacgtat atcgaaagtc
     1561 tggcagctgc gctgccagat gaaattctta ctagtgcaga gatcgagcgc cggctggcgc
     1621 cgctgtatga acgcctgcac ctgcccgaag gtcgcctcga actgatgacc ggcattaggg
     1681 agagacgtct gtggcctgca ggtactcgtg cttcagatgc ctctgctcgg gcgggtagag
     1741 ccgcactggc gaaatcatgc attcggcctg aacagattga gctgtacatc catgcggctg
     1801 tgtgtcggga tatgctggaa ccagccacag cggcatttgc tcacaggaaa atcggcttat
     1861 caccgaactg ccagattttt gacttgtcca atgcgtgcct cgggttcctc aatgcattaa
     1921 ctcttgctgc atctatgatc gaggcgggac agattcgcgc cgcattaatt acttgcggcg
     1981 aagattccag accacttgtt gaacagaccc tccgcactct gaacgccgag ccactgacgc
     2041 gtaatggtat caaaccctat tttgccaatc tgaccgtagg gtcaggggcg gtgggcgctg
     2101 tcgtctgcca cgaaagtctt gtggttccgg agggtctggg cagtggtggt gccgcaggcg
     2161 aatcgagccg tgctcaccgc cttatcggtg gggtggtccg tgccgcaaca gcccatagtg
     2221 agctttgcca gggcgaaacg catggggcag atagcctcgc catgcagaca gacagcgagg
     2281 ccttactaat agccggagtt actttggcgc acgaaacctg gggtcagttc tgcgaagaaa
     2341 ccggatggga cgcagctacc ccgtctcgtt ttatttgcca tcaagtcggt tcgatccatc
     2401 ggcgcaaact ttacgatacg ttaggtatag atttggctaa ggacttttcg actttcgagt
     2461 ttttgggaaa cactgggtca gtagcactcc cagcgacatt agcacttgcg gtcgaaaaag
     2521 gtgttgtgcg ctctggtgac aaggtagcat tacttggtat tggcagtggt ctgaattcac
     2581 tgatgttagc tgtagaatgg tgactcgagc accaccacca ccaccactga gatccggctg
     2641 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2701 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     2761 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2821 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2881 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2941 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3001 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3061 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3121 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3181 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3301 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     3361 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3421 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3481 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3541 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3601 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3661 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3721 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3781 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3841 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3901 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3961 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     4021 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     4081 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     4141 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     4201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4381 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     5041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     5101 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     5161 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5221 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5281 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     5341 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     5401 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     5461 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     5521 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     5581 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     5641 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     5701 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     5761 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     5821 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     5881 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     5941 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     6001 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     6061 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     6121 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     6181 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     6241 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6301 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6361 att
//
