LOCUS       sequence-317817-        6309 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 6309)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6309)
  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_017959
FEATURES             Location/Qualifiers
     source          1..6309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     893..912
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(932..1009)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1215..1234
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1318..1337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1318..1336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1337..1361
                     /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             1376..1398
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1417..1434
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1444..1461
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2491..2508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2561..2579)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2575..2622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2659..3114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2746..2765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2956..2977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3206..4021)
                     /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(3206..4021)
                     /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     3929..3948
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4013..4032
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4143..4731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4632..4651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4885..4902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4917..5056
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4982..5001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5142..5164
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5158..5349)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6158..6309,1..931))
                     /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"
ORIGIN
        1 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
       61 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
      121 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
      181 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
      241 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
      301 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
      361 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
      421 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
      481 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
      541 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
      601 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
      661 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
      721 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
      781 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
      841 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
      901 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
      961 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     1021 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     1081 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     1141 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     1201 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     1261 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     1321 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     1381 ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca tcacagcagc
     1441 ggcctggtgc cgcgcggcag ccatatgtct gaaaacgcga gctatagact cgcgaattgt
     1501 tcaatacttt cggtaagtac tgtcgaagca ccggaagtag ttaccagcga ccatcttgac
     1561 gaattacttc aaccggcgta cgatacgaca gggatgatgg cagggcagat agaacggtta
     1621 gcaggagtgc gagaacgcag gtggtttcct aaagatgctg attatactcg cggcgcggtg
     1681 gaggcggggc gtcaagccat agctgatgca cgtattgatg ccgatcggat tggtttaatt
     1741 gttaacgcaa gtgtgacccg tccgcatctc gaacccggta tctcggcaat ggttcatgca
     1801 gagctcgaac tgccacgttc atgtatagct tttgatgtca ccaatgcctg cctgggggta
     1861 gttaacagtc ttcaaatcgc tggcactatg atagatgcag ggcagattga atacgcactg
     1921 atagtagcca gcgagggcgc ccgacagatg caggaagcca gcatcgctcg tttggtggat
     1981 ggtggcagtc gccaagatgt caaagatgcc tttgccacga tgacgctggg tagcggcgcc
     2041 gtgggaattg ttctgggccg aagtgatcga catccggagg gacatcagat tattggtggt
     2101 atcacacgcg ctggcactga gcaccatgag ctgtgtatag gtgggatgga tggaatgcgt
     2161 actgatgcga gcaagctgtt tgtagaaggg ctgagtttag cgaccgatgc ttgggctgaa
     2221 gcaaaagggg agtgggattg ggaagaaatg gattggtacg tggcccatca gacctccact
     2281 gcgcacatcg cctgtctgtg cgatacgctg aagctccctg cggaaaaatt cccggttact
     2341 gttgatacgt atggcaatat cggtccggtt gcactcccat tcacgctcgg aatgtatcag
     2401 cacaaaatga gaaaagggga tcgcgtgctg cttatgggta taggtagcgg tctgaatacc
     2461 agcatgactg aggtgcgctg gtaactcgag caccaccacc accaccactg agatccggct
     2521 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2581 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2641 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2701 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2761 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2821 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2881 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2941 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3001 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3061 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3181 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     3241 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3301 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3361 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3421 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     3481 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3541 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     3601 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3661 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     3721 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3781 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     3841 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     3901 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     3961 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     4021 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     4081 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4141 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4201 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4261 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4321 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4381 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4441 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4501 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4561 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4621 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4681 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4741 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4801 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4861 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4921 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     4981 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     5041 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5101 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5161 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5221 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5281 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5341 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     5401 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     5461 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     5521 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     5581 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     5641 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     5701 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     5761 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     5821 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     5881 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     5941 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     6001 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     6061 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     6121 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6181 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6241 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     6301 tcaccgcct
//
