LOCUS       sequence-317452-        6312 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 6312)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6312)
  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_017965
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..128
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    231..255
                     /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             270..292
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             311..328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             338..355
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1388..1405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1458..1476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1472..1519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1556..2011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1643..1662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1853..1874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2103..2918)
                     /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(2103..2918)
                     /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     2826..2845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2910..2929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3040..3628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3529..3548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3782..3799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3814..3953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3879..3898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4039..4061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4055..4246)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5055..6137)
                     /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     6099..6118
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6138..6215)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
       61 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      121 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      181 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      241 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
      301 gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat
      361 ggatgggaac gcaacgacac gctttagtaa tgtatcactg ttggccgtaa ctcgccgtct
      421 gcccagccgg gtaaccacgt cggcagagat tcagggtctg ctggcaccgg cgcttcggcg
      481 tcttcgcgta ccgggccggc ttttagagcg agtggcgggg gtccatgccc ggagatcatg
      541 gggacctgac gaagattgcg acgccgcgac ggtcgcagca gcacaagaag ctctggaccg
      601 cgctggagta gcaccagacg atgtggggtt gctggttaat acatctgtga cgcgcaaaca
      661 tttagaaccg agcgtggcag tcgccctgca tcacggcctc ggcctgccgt cgagtgccgt
      721 aaatttcgat gtggcaaatg cgtgtctcgg gtttataaac ggtatgtcga ttgcagctgg
      781 tatgattgag tccggtcaga tacgttatgc gttggttgtg gatggtgagg atgctgatga
      841 ggtgcgtgcg accacggttg aacgtctgct tagagatggt gtgggccgcg aagacttcat
      901 gcaggagttc gcaacgctga cgctgggctc cggaagtgcg accgctgtac ttggtagagc
      961 tgacgaacac ccggaaggac atcgcctctt gggtggggta actcgtgccg ccactcgctt
     1021 tcacgacctt tgcgtgggat cagcgaaagg aatgtttaca gacgcacgcg cactgctcga
     1081 cggcgggctc gaacttgttg tggacgcgtg gaaagaggct gtaagtgaat gggattgggc
     1141 atcaatggat cgttatgtga ctcatcaagt ctccagggtc catacagacg ccatcgtcga
     1201 agctgtggga ctggaccgta gtaaggttcc gaccacattc ccccgtttgg ggaatgtggg
     1261 ccccgcaagc gtcccgatta cgctagctga agaagctgaa acgctccgac ggggtgatag
     1321 ggttttattg atgggagtag gatccggtat caacaccgca atgatggaac tggcgtggta
     1381 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1441 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1501 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1561 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1621 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1681 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1741 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1801 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1861 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1921 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1981 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     2041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     2101 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2161 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2221 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2281 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2341 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2401 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2461 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2521 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2581 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2641 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2701 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2761 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2821 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2881 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2941 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     3001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3181 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3781 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3841 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3901 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3961 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4021 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4081 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4141 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4201 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4261 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4321 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4381 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4441 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4501 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4561 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4621 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4681 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     4741 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4801 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4861 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4921 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4981 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     5041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     5161 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     5221 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     5281 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     5341 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     5401 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     5461 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     5521 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     5581 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     5641 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     5701 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     5761 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     5821 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     5881 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     5941 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     6001 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     6061 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     6121 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     6181 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     6241 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     6301 aaggaatggt gc
//
