LOCUS       sequence-317816-        6558 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 6558)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6558)
  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_018095
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     271..290
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      401..989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     890..909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1143..1160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1175..1314
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1240..1259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1400..1422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1416..1607)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2416..3498)
                     /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     3460..3479
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3499..3576)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3782..3801
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3885..3904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3885..3903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3904..3928
                     /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             3943..3965
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3984..4001
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4011..4028
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5061..5078
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5131..5149)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5145..5192
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5229..5684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5316..5335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5526..5547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
       61 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
      121 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
      181 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
      241 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
      301 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
      361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
      421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
      481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
      541 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
      601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1141 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1201 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     1261 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     1321 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     1381 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1441 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     1501 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     1561 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     1621 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     1681 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     1741 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     1801 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     1861 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     1921 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     1981 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     2041 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     2101 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     2161 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     2221 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     2281 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     2341 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     2401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     2521 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     2581 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     2641 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     2701 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     2761 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     2821 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     2881 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     2941 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     3001 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     3061 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     3121 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     3181 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     3241 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     3301 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     3361 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     3421 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     3481 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     3541 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     3601 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     3661 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     3721 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     3781 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     3841 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     3901 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
     3961 ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc
     4021 gcggcagcca tatgacaggt aatagcgact ttcgatttca tgacgtggcg attctgagtc
     4081 tggacgcagg gccggccccg ttagtagtca cttcagcaga tgtggatcgc aggctggccc
     4141 cgttttacgc aagaactcgt agcatgagtg gtttagtgac ttccctggcc ggtgattttg
     4201 aaaggcgtca atggccggca gaagtcagct ttattcaagc agcagcccgc gcaggagaag
     4261 ctgctatagc agcagcaggc attgataggg ctaggattgg tgttgtgatt gatacctcag
     4321 tttcacgcgc tcgtctggag ccatcctctg cggtctcggt gcatcattta cttgatttac
     4381 ctccgaactg cttaaatttt gatgtaagca atgcgtgtct gggtttcgtt acgggtctgc
     4441 atctggccgg agcattaatc gattcaggcc aaattgatta tgctttgatc gttgatggcg
     4501 aaggcaccag agaaatctac gatcgtacaa ttgatcgttt aaatgaacgt ggtgaaggtc
     4561 ttcctgatct gctggccaac tttgctacct ttactctggg atctggtgct gctgccatgg
     4621 ttctggggcg gcactctgcc cacccggagg gtcatcagtt attagatggc ttttttcgcg
     4681 cggaaacacg gcatcacgag ctgtgtattg gtacaatcga cggcggcgtg acggactcac
     4741 gtgctctcct ggaggcggga attgaattag caggcgctgc atgggcacag gctggagcaa
     4801 ctgcagcttg gggctctcgt gatagatata ttttccacca agtgtctgag gttcacacga
     4861 aagctatgat ccgcactctg ggcgtagacc ccgcaaaagt tcctaccacc ttcgcgagat
     4921 acggcaatat tggtccggcc gctattccca ttacacttgc aacggttgcc gatgaactca
     4981 gcccaggcga tattgtactc tgtatgggta ttgggtccgg cttaaatgtg ggtgtgattg
     5041 agctgcgctg gtgactcgag caccaccacc accaccactg agatccggct gctaacaaag
     5101 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5161 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     5221 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5281 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5341 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5401 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5461 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5521 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5581 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5641 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5761 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     5821 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     5881 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     5941 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     6001 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     6061 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     6121 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6181 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     6241 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6301 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6361 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6421 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6481 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6541 ggaatttaat cgcggcct
//
