LOCUS       sequence-316707-        6360 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 6360)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6360)
  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_018151
FEATURES             Location/Qualifiers
     source          1..6360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             56..73
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             83..100
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1181..1198
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1251..1269)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1265..1312
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1349..1804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1436..1455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1646..1667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1896..2711)
                     /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(1896..2711)
                     /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     2619..2638
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2703..2722
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2833..3421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3322..3341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3575..3592
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3607..3746
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3672..3691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3832..3854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3848..4039)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4848..5930)
                     /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     5892..5911
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5931..6008)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6214..6233
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6317..6336
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6317..6335
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6336..6360
                     /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)."
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat accatgggca gcagccatca
       61 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatgtcgg cgcccagtcc
      121 aacaccgcgg cgcctggtac tggtcacaaa cgtttctggt aatgccagtt atgaatattc
      181 tcgcgccgcg ctgctggctg ttacagaagt agaggcaccg gtggaggtta catcgcagga
      241 cctggatgat caattagccg aagttcttac cagactccgc ttgccgggtt cgcttctgga
      301 acgtgtcgcc ggagtccgct ctagacgtaa ctgggatgag accgaagatt ttcaggaggc
      361 ggcagcgcga gcaggagtag cggcgctaga acaagcgggt gttcctagag aacgggtagg
      421 tctacttgta aatacctctg tgacccgagc gacgttagaa ccttctgtag ccgtccgtat
      481 ccatcatctt atgggtctgc ctacctcagc aacaaatttt gatattacaa atgcatgcct
      541 gggtttcgta aatggtctca tgttagcatc cacccttgtt gacgctggcc aaattgatta
      601 tgcggtagtt gtggctgcag aagatgcaag caaagtacag gccgccaccg tagcgaactt
      661 gcatgctgaa accacacacc gtggtaattt catggaacag tttgctagct tgactcttgg
      721 aagtggcgct gcggctgcag tagttggtcg cattgaagac catccggatg cacatcgtat
      781 tgtacgcgga atcacccgcg ccggcacaga ccatcacggc ctatgtgtag gcgatcacca
      841 tggtatgtat accgacagta ctgccctcct caaagatggc ttagaattgg tcatggatgc
      901 ctggcatgat gtacctgcgg agtgggcgtg gcctgaagct gatcggttta ttactcatca
      961 agtgagccag atgcataccg atgcaattgc cgaatcagcg catcttgatc cagcacgtat
     1021 acctacgacc tttccaactc tgggaaatgt gggacccgca tcattgccta tcactctggt
     1081 tagggaaatg gacgctcttg aggatgggga tactgtggtg tgcctgggcg tcggtagcgg
     1141 attgaacaca gcgatgctgg agatcgagtg gtagctcgag caccaccacc accaccactg
     1201 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1261 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1321 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1381 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1441 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1501 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1561 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1621 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1681 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1741 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1801 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1861 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1921 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1981 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2041 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2101 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2161 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2221 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2281 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2341 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2401 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2461 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2521 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2581 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2641 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2701 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3481 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3541 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3601 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     3661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3721 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3781 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3841 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3901 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3961 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4021 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4081 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4141 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4201 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4261 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4321 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4381 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4441 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     4501 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     4561 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     4621 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     4681 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     4741 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4801 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4861 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4921 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4981 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     5041 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     5101 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     5161 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     5221 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     5281 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5341 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5401 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5461 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5521 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5581 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5641 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5701 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5761 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5821 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5881 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5941 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     6001 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     6061 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     6121 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6181 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     6241 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     6301 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
//
