LOCUS       sequence-318293-        6306 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 6306)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6306)
  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_017957
FEATURES             Location/Qualifiers
     source          1..6306
                     /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             2488..2505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2558..2576)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2572..2619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2656..3111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2953..2974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3203..4018)
                     /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(3203..4018)
                     /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     3926..3945
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4010..4029
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4140..4728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4629..4648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4882..4899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4914..5053
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4979..4998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5139..5161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5155..5346)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6155..6306,1..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 ccatatgctg tttaaaaatg tagcgattgc cggtcttgca
     1501 catgtggatg cccctcattc cctgacttcc gatgaaatta atctgcgtct gaaacccacg
     1561 ctggatagac taggtatccg cgttgatgtg cttaaagata ttgctggaat acacgcccgc
     1621 agattgtggg gatcagataa acttgcctcg gacgcggcga cggaagcagg cattcttgcc
     1681 ctaaaggatg ccggcattca aagcggccaa ctcggtttat tagtttccac cagcgtttcg
     1741 cgtgactatc tggagccgag caccgcaagt atcgtagcag gcaatctgca ggttggtgac
     1801 acatgccaga attatgacgt tgcaaatgcc tgcttggctt ttatcaatgg tatggatatg
     1861 gcagcacgca tgatagagag aggtgagatt gattatgcgc tagtcgttga cggtgaaaca
     1921 gccgatttgg cttatgagaa aactattgag agaatgctga ggccggactc gacagaagat
     1981 gaatttcgca atgaactggc tacgctgacg ctgggctctg gggcggcagg tatggtactg
     2041 gcccgcgctg agcttcatcc ggaagcgcct cgctacaagg ggggcgtcac ccgttcggcc
     2101 acggaatgga atacgttgtg cagaggtaac ctggatcgga tggtgactga cacgaaacag
     2161 ctcctggttc agggaatgaa tctggccggg aaaactttcg tagctgccaa acaggctatg
     2221 ggttgggctg ttcatgaact ggatcagttt gtggtacatc aggtgagcaa agtacataca
     2281 cagacactga tccaaaaatt tgggattgat cccaaaaagg taatgacgat ttttggtgag
     2341 catggtaata taggccctgc ctcggtaccg attgtactgt ccaagctgaa acaggccggg
     2401 cgactgaaaa agggagatcg tattgccctg ctggggattg gctcaggttt aaactgcaca
     2461 atgtcagaag ttgtgtggta gctcgagcac caccaccacc accactgaga tccggctgct
     2521 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2581 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2641 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3001 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3061 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     3241 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     3301 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     3361 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     3421 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     3481 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     3541 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     3601 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     3661 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     3721 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     3781 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     3841 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     3901 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     3961 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     4021 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     4081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4261 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4921 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4981 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     5041 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5101 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5161 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5221 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5281 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5341 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     5401 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     5461 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5521 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5581 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5641 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5701 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5761 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     5821 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     5881 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     5941 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     6001 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     6061 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     6121 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6241 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     6301 ccgcct
//
