LOCUS       sequence-315778-        6318 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Hydrocarbon Structures for Fuels and Chemicals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6318)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: JGI_018129
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     719..736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    751..890
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(816..835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     976..998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(992..1183)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1992..3074)
                     /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     3036..3055
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3075..3152)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3358..3377
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3461..3480
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3461..3479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3480..3504
                     /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             3519..3541
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3560..3577
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3587..3604
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4643..4660
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4713..4731)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4727..4774
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4811..5266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4898..4917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5108..5129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5358..6173)
                     /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(5358..6173)
                     /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     6081..6100
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6165..6184
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6295..6318,1..565)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
       61 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      121 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      661 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      721 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
      781 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
      841 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
      901 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      961 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1021 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     1081 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     1141 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     1201 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     1261 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     1321 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     1381 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     1441 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     1501 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     1561 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     1621 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct
     1681 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     1741 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     1801 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     1861 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     1921 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     1981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     2101 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     2161 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     2221 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     2281 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     2341 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     2401 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     2461 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     2521 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     2581 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     2641 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     2701 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     2761 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     2821 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     2881 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     2941 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     3001 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     3061 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     3121 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     3181 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     3241 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     3301 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     3361 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     3421 gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg
     3481 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag
     3541 atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcctgg tgccgcgcgg
     3601 cagccatatg cggaacgacg cccctccgca tggtaatgaa attccaaaag aacagccgtt
     3661 cgcgaacccg caaagtcaga ccaatgagac cccggttcac agtccagaga gtggtaaccc
     3721 acattccgca atacgtaacc cgcatagttc tcgtccagac tggcttcgtg ctctataccc
     3781 gttcgcacaa aacacattcg taaccccgcg tggtgcacgt atgaactatg ccgatgaggg
     3841 tccgcgtact gacgaagcgg tgctgatgct gcatggtaac cccacttggt cttttttcta
     3901 tcgcgatatc atacgcgcgc tttcaccgtc tctgcgctgc attgcaccgg atcatattgg
     3961 aatgggcctg tctgagaaac cgtctgcccg tgattatcca tatacactgg cacaacgtat
     4021 agctgacatc gagaccctgg tggcctccct gaatttacga cggatccatc tgattgtcca
     4081 tgattggggg ggcgccattg gctttggttt cgcgacccga caccccgaga aaattggccg
     4141 cattactata ctgaacaccg cagcttttcc ggacatccac atgccggctc gtattggctt
     4201 atgccgtgcc cccttaggca tcggtgctct catcgttcgg gggttcaacg gctttgcatg
     4261 gccggctacg tggatgacga tgcaccgacg taaactgacc ccagatgaaa agagagctta
     4321 cctttttccc cataacaact gggccaatcg cgtggcagta cacgagtttg tgcgggatat
     4381 cccgatgtca cctgctcatc caacccacgc tactctgacc gccatcgaac gtaatttacc
     4441 gttgctggtt gcgaatccaa agttaattgt atggggaggc gcggattttt gctttaatga
     4501 ccacttttat aatcggtggc gaaaagtcta tccagaagct gcaacccacc actttcccga
     4561 tgtagggcat tacctactgg atgatggcgg cgaagaagtt cgcgcgaaaa tcgttcaatt
     4621 ctttacgcag tcttaactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     4681 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4741 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     4801 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4861 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4921 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4981 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5041 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5101 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5161 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5221 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     5281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5341 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     5401 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     5461 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     5521 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     5581 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     5641 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     5701 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     5761 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     5821 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     5881 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     5941 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     6001 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     6061 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     6121 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     6181 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     6241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6301 tccttttttt ctgcgcgt
//
