LOCUS       sequence-317809-        6228 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 6228)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6228)
  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_017931
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     940..959
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(979..1056)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1262..1281
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1365..1384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1365..1383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1384..1408
                     /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             1423..1445
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1464..1481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1491..1508
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2457..2474
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2527..2545)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2541..2588
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2625..3080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2712..2731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2922..2943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3172..3987)
                     /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(3172..3987)
                     /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     3895..3914
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3979..3998
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4851..4868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4883..5022
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4948..4967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5108..5130
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5124..5315)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6124..6228,1..978))
                     /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 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
       61 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
      121 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
      181 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
      241 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
      301 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
      361 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
      421 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
      481 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
      541 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
      601 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
      661 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
      721 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
      781 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
      841 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
      901 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
      961 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     1021 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     1081 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     1141 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     1201 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     1261 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     1321 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     1381 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
     1441 ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc
     1501 gcggcagcca tatgaatgtg ggcctgctgg gtacgggtca ttacgttccg acaaatgtgg
     1561 tgacaaataa ggatatggaa aaaattgttg atacgaatga tgagtggatc agaacccgca
     1621 cgggtattga agagcggaga attgcatctg atgatattaa tacctcagat atggcatttt
     1681 ttgcggctca agaagccctt aaggaatcta atgtgaaagc ggaagatata gatttgatcc
     1741 tggttgcaac tgttacacca gacacgccat tcccgagcgt cagctgtatg attcaggacc
     1801 gccttggtgc acgtaaagca gcagctatgg atatctctgc ggcttgtagc ggttttatgt
     1861 atggagtagt cacagccaaa cagttcattg agactaatgc ctacaaaaac atattggtag
     1921 tcggcgcgga aaaactcagc aaaataacag attggtccga tcgtaatacc tgtgtactgt
     1981 ttggagatgg agctggagct gcagtggtcg ggccagtctc ggagggaaaa ggcattctga
     2041 gttttgaact cggcgcggac ggcagcggcg ggaaagaatt atatcagaac gacgaagacc
     2101 atctcatgat gaacggtagg gaagttttta aatttgccgt gcggcaaatg ccagatagtt
     2161 cagtgaacgt cgttgaacaa gcaggctaca acaaagagga tgttgattat ctagtcccac
     2221 atcaggctaa tatacgtatc atggaagccg cacgcgaaaa gttaggtatc agcgaagaaa
     2281 agatggcgac tagcattaaa cgttatggca ataatagtag cgccagtata ccgattgcac
     2341 tgtcggaggg ggttaaaagc ggcaaaatta aagatgatga tttggttgtt ctggttggct
     2401 tcgggggtgg gcttacgtgg ggtgccatcg ccttgcgctg gggcaaatga ctcgagcacc
     2461 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     2521 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2581 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     2641 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2701 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2761 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2821 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2881 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2941 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3001 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3061 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     3181 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3241 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3301 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3361 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3421 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3481 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3541 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3601 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3661 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3721 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3781 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3841 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3901 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     3961 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     4021 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4801 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4861 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4921 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4981 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     5041 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5101 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     5161 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     5221 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     5281 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5341 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5401 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5461 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5521 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5581 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5641 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5701 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5761 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5821 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5881 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5941 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     6001 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     6061 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     6121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttt
//
