LOCUS       sequence-317184-        6330 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 6330)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6330)
  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_017983
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1341..1532
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1526..1548)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1634..1773
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1689..1708
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1788..1805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2658..2677)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2669..3484
                     /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             2669..3484
                     /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     complement(2742..2761)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3576..4031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3713..3734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3925..3944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4068..4115)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4111..4129
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4182..4199)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5250..5267)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(5277..5294)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5313..5335
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5350..5374
                     /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)."
     primer_bind     complement(5374..5393)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5375..5393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5477..5496)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5702..5779
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5780..6330,1..532)
                     /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     complement(5799..5818)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
       61 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
      121 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
      181 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
      241 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
      301 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
      361 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
      421 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
      481 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      541 caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag
      601 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
      661 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
      721 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
      781 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
      841 agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt
      901 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
      961 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     1021 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     1081 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     1141 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     1201 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     1261 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     1321 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     1381 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     1441 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     1501 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1561 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1621 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     1681 taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg
     1741 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     1801 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1861 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc
     2641 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     2701 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2761 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     2821 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2881 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2941 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     3001 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3061 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3121 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3181 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3241 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3301 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3361 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3421 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3481 ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3541 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     3601 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3661 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3721 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3781 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3841 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3901 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3961 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4021 acagggcgcg tcccattcgc caatccggat atagttcctc ctttcagcaa aaaacccctc
     4081 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
     4141 aactcagctt cctttcgggc tttgttagca gccggatctc agtggtggtg gtggtggtgc
     4201 tcgagtcacc agtcaatacc caacatcagt gcattaatgc cactcccgat acccaggatg
     4261 gctgctcgtt ggccaggcac aagtgcccct tgttcaatgc ccagcgccat cgtgatagga
     4321 gctgagacgc tcccgacatt ccctaatgtt ggcagggttt caaaatttct agcgggatct
     4381 aactgcaatg tttcaaacag catctgcgcg tgtgctttgc caacctggtg acaaaaaaac
     4441 ctgtcaatgg cacccttgtc ccaaccaagc gcagtctgga acgaccgcca acatgcggct
     4501 gcggtctcaa tgcctttttc tagcaactgt tcggaatcgg ttgacattaa tgtaccttgt
     4561 tctgcgttct ggccgccctg acataaatcg ttatgggccg tattggcgcg acacgcgcct
     4621 cccctaaggt aatggccagt gtcacggcaa tcggaacgcg acattacaag agcaacagca
     4681 ccgcttccta tggtcaaact tgcgaacagg ggtttaatag ttttacgggt tagcgttgta
     4741 tctgccagca gatgagcaag agtggaatcg atcaagttct cagcggtctc tccggccacc
     4801 actaaacccg cccgcacttg gcccatttcc agcatatttg ccagcatcac cattccatca
     4861 agaaacccta aacaagcatt gctaatgtcg aacagaaggc aatctggcgg taatccaagc
     4921 gcgcggtgca caaaggccgc agtagcaggt tccatcatat ctcgggatac ggacgtaaag
     4981 agtaaacact ccagttccgc gacatcaatg cctgcacgtg cgatagcagc agcccctgct
     5041 gcggctgccc ctgcactcgg acgtgtgccc ggctgccaca gacgacgggc agcaataccg
     5101 gtcatcagct ccaggcggcc ggccggaagc ttcaggcgct cgtacaaggg tcctaaacgc
     5161 tcttcaatgg tggtacttga taattctacc ggaggcagtt gatacccaaa ttcatgcaag
     5221 cagactctac tatatcttaa attcatatgg ctgccgcgcg gcaccaggcc gctgctgtga
     5281 tgatgatgat gatggctgct gcccatggta tatctccttc ttaaagttaa acaaaattat
     5341 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc
     5401 gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     5461 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     5521 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     5581 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     5641 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc
     5701 ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     5761 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     5821 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     5881 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     5941 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     6001 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     6061 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     6121 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     6181 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     6241 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     6301 ggtcgcattg ggtcaccagc aaatcgcgct
//
