LOCUS       sequence-317631-        6297 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 6297)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6297)
  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_017949
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1162..1179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1232..1250)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1246..1293
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1330..1785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1417..1436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1627..1648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1877..2692)
                     /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(1877..2692)
                     /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     2600..2619
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2684..2703
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2814..3402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3303..3322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3556..3573
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3588..3727
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3653..3672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3813..3835
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3829..4020)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4829..5911)
                     /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     5873..5892
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5912..5989)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6195..6214
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg acctattctc gtatcggtca aatcgcagtc
      181 catctgccag aaggaggacg taccagcgcg gaggtcgaag cgctggtggc cgagctgaac
      241 ccgggtttcg aacctcccga gggcttgata gaacgcttta caggagtgcg ttatcgccat
      301 attgccccgg aagacacaca agcttctgat ttagctgttg cggctatcga gaagctgttt
      361 gcgtcatggg gacatggtat tgatgacgtt gatctgctga tttttgccgc ggcctctacg
      421 gatgttattg aaccggcgac agcccatatc gtctccgaca aactgggcgg tagttgtcca
      481 gtattcgata ttagcaatgc gtgttcgagt gtcttcaatg ctatagaagt agcggatgct
      541 ctgatccgca cgggccagta tcgtagagtg ttgatcgcgt gtggcgaacg agtgagtcct
      601 ggagcaaggt accgtgatgt tgcaacaatg gatgactttg tgcatgcggg agcaagccat
      661 acagtttcag atgcgggcgc tgccctgctc gttgaagcca gtgatgagcc gggtattata
      721 ggacatcgtt caatagcgaa ctctagtgcc tggtctgcaa ctgtagtgcc actggtgcat
      781 gataaagagt cagacacact taaagtcgaa tatttgggtg ttaaaacgtt ggacctgatg
      841 aaaggacttg atcaggcgga tattacgcct ttgttagatg cggttgctgg ctttggatta
      901 actatggatg actttgacgt tgtttgtatc catcaggcgg tgcttgctta tgtggaagta
      961 ttttgtgaac gcgcaggcat tccgtacgat cgaacagttt taacactggc cgaccacggc
     1021 aacgttgcct ctgctacact gcctcttcaa ctggtgaagg cagttgagag cggacgtctt
     1081 aatccgggcg acaaagtgct gttagtagga ttggcatctg ggctgagctt gggtgttatg
     1141 gctatgcagt ggtaactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     1201 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     1261 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     1321 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1381 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1441 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1501 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1561 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1621 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1681 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1741 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1861 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1921 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1981 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2041 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2101 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     2161 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2221 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2281 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     2341 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2401 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2461 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2521 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2581 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2641 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     2701 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     2761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2941 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3481 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3541 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3601 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     3661 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3721 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3781 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3841 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3901 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     3961 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4021 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4081 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4141 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4201 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     4261 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4321 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4381 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     4441 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     4501 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     4561 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     4621 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     4681 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4741 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     4801 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4981 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5041 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5101 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     5161 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5221 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5281 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5341 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5401 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5461 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5521 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5581 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5641 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5701 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5761 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5821 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5881 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5941 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     6001 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6061 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6121 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6181 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6241 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
