LOCUS       sequence-316880-        6315 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 6315)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6315)
  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_018103
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      48..636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     537..556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     790..807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    822..961
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(887..906)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1047..1069
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1063..1254)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2063..3145)
                     /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     3107..3126
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3146..3223)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3429..3448
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3532..3551
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3532..3550
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3551..3575
                     /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             3590..3612
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3631..3648
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3658..3675
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4711..4728
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4781..4799)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4795..4842
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4879..5334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4966..4985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5176..5197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5426..6241)
                     /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(5426..6241)
                     /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     6149..6168
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6233..6252
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
       61 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      181 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      721 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      781 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
      841 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
      901 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
      961 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     1021 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     1081 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     1141 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     1201 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     1261 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     1321 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     1381 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     1441 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     1501 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     1561 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     1621 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     1681 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     1741 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     1801 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     1861 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     1921 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     1981 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     2041 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     2161 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     2221 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     2281 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     2341 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     2401 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     2461 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     2521 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     2581 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     2641 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     2701 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     2761 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     2821 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     2881 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     2941 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     3001 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     3061 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     3121 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     3181 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     3241 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     3301 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     3361 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     3421 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     3481 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     3541 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     3601 ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg
     3661 gtgccgcgcg gcagccatat gagcggaaat gccacccacc ggttcggaaa cacggccgtt
     3721 ctggctttac aaaccacaga tgcaacaaga gttgtgacat ctgacgctct tgatgatgcg
     3781 ctagcagata cgtatcgtcg ggtaggtctt cgcccgggac ttctggaacg actcgccgga
     3841 attcgtgaaa gacgctggtg gggagaaggt gtgacttttg tagatggcgc ggccgaagcc
     3901 ggtgcaaaag caatctcgga aagtggtgta gatcctgccg gtattggcgt catgattaat
     3961 acgagcgtat cgcgtaggtg gctggagcct agtacagccg tggccgttca ccatgctctc
     4021 ggactacccc gtagctgtca gaattttgac gtaaccaatg cttgtcttgg ttttgtcaac
     4081 gggatggaac tagctgctgc catgatcgac gccggcttgg tggaacatgc tcttgtggtc
     4141 aatggtgaag atgcacagcc ggtacaggag cggaccctgg atcgattaaa ccagcccggt
     4201 accggctcaa aagatgtcat tgcgcagttt gcaacactaa ctttaggttc tggcgcggct
     4261 gctatggtat tgggtcgcgc tgaccgtcat ccggaaggtc atagactggt cgcgtctgtg
     4321 tcacgcgcgg gtaccgaaca tcacgagctg tgtgttgggg acaacgatct gatgcggacg
     4381 gacttaaaag gtctgctgga tgccgggctg gcgttatcag aagatatgtg ggcagatgcg
     4441 gcagccgaag tcgattgggc tggcggaatg tcacgttata tcgttcacca ggtgtctaaa
     4501 gtacacacgc aggcgatgtg tgatagattc tcgatagatc cagcgttagt tccgacgacg
     4561 tttcctacac gcggcaattt gggtccagcc agcgttccgt acactctggc tggagaacag
     4621 gatagtttag tggatgggga tcgggtactt ctgatgggta ttggttcggg gctgaatgtc
     4681 tcctgcctag aactggcatg gtaactcgag caccaccacc accaccactg agatccggct
     4741 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     4801 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     4861 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4921 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4981 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5041 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5101 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5161 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5221 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5281 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     5341 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5401 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     5461 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     5521 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     5581 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     5641 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     5701 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     5761 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     5821 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     5881 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     5941 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     6001 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     6061 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     6121 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     6181 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     6241 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     6301 ccttaacgtg agttt
//
