LOCUS       sequence-367933-        6399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CDS of D-HicDH codon optimized for expression in Synechocystis sp. 
            PCC 6803 under the control of the synthetic promoter Ptrc.x.tetO2 
            and the RBS BBa_B0030 in pSEVA251 plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  AUTHORS   Jurkas V, Winkler CK, Poschenrieder S, Oliveira P, Pacheco CC, 
            Ferreira EA, Weissensteiner F, De Santis P, Kara S, Kourist R, 
            Tamagnini P, Kroutil W
  TITLE     Expression and activity of heterologous hydroxyisocaproate 
            dehydrogenases in Synechocystis sp. PCC 6803 DeltahoxYH.
  JOURNAL   Eng Microbiol. 2021 Nov 26;2(1):100008. doi: 
            10.1016/j.engmic.2021.100008. eCollection 2022 Mar.
  PUBMED    39628613
REFERENCE   2  (bases 1 to 6399)
  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"; journalName: "Eng 
            Microbiol."; date: "2021-11-26"; volume: "2(1):100008. doi"; pages: 
            " 10.1016/j.engmic.2021.100008. eCollection 2022 Mar"
COMMENT     Sequence Label: pSEVA251_ Ptrc.x.tetO2::D-HicDH
FEATURES             Location/Qualifiers
     source          1..6399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(968..1939)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     rep_origin      2046..2440
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(2478..2564)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2579..2601
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    2626..2647
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        join(2653..2658,2659..2675,2676..2682)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     RBS             2727..2738
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2748..2765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3777..3797
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3852..3946
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4067..4086)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4078..4893
                     /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"
     oriT            5060..5168
                     /note="incP origin of transfer"
     CDS             complement(5179..6030)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(join(6017..6399,1..457))
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
ORIGIN
        1 tggcgttttc ttcctcgatg tggaaccggc gcagcgtgtc cagcaccatc aggcggcggc
       61 cctcggcggc gcgcttgagg ccgtcgaacc actccggggc catgatgttg ggcaggctgc
      121 cgatcagcgg ctggatcagc aggccgtcag ccacggcttg ccgttcctcg gcgctgaggt
      181 gcgccccaag ggcgtgcagg cggtgatgaa tggcggtggg cgggtcttcg gcgggcaggt
      241 agatcaccgg gccggtgggc agttcgccca cctccagcag atccggcccg cctgcaatct
      301 gtgcggccag ttgcagggcc agcatggatt taccggcacc accgggcgac accagcgccc
      361 cgaccgtacc ggccaccatg ttgggcaaaa cgtagtccag cggtggcggc gctgctgcga
      421 acgcctccag aatattgata ggcttatggg tagccattga ttgcctcctt tgcaggcagt
      481 tggtggttag gcgctggcgg ggtcactacc cccgccctgc gccgctctga gttcttccag
      541 gcactcgcgc agcgcctcgt attcgtcgtc ggtcagccag aacttgcgct gacgcatccc
      601 tttggccttc atgcgctcgg catatcgcgc ttggcgtaca gcgtcagggc tggccagcag
      661 gtcgccggtc tgcttgtcct tttggtcttt catatcagtc accgagaaac ttgccggggc
      721 cgaaaggctt gtcttcgcgg aacaaggaca aggtgcagcc gtcaaggtta aggctggcca
      781 tatcagcgac tgaaaagcgg ccagcctcgg ccttgtttga cgtataacca aagccaccgg
      841 gcaaccaata gcccttgtca cttttgatca ggtagaccga ccctgaagcg cttttttcgt
      901 attccataaa acccccttct gtgcgtgagt actcatagta taacaggcgt gagtaccaac
      961 gcaagcacta catgctgaaa tctggcccgc ccctgtccat gcctcgctgg cggggtgccg
     1021 gtgcccgtgc cagctcggcc cgcgcaagct ggacgctggg cagacccatg accttgctga
     1081 cggtgcgctc gatgtaatcc gcttcgtggc cgggcttgcg ctctgccagc gctgggctgg
     1141 cctcggccat ggccttgccg atttcctcgg cactgcggcc ccggctggcc agcttctgcg
     1201 cggcgataaa gtcgcacttg ctgaggtcat caccgaagcg cttgaccagc ccggccatct
     1261 cgctgcggta ctcgtccagc gccgtgcgcc ggtggcggct aagctgccgc tcgggcagtt
     1321 cgaggctggc cagcctgcgg gccttctcct gctgccgctg ggcctgctcg atctgctggc
     1381 cagcctgctg caccagcgcc gggccagcgg tggcggtctt gcccttggat tcacgcagca
     1441 gcacccacgg ctgataaccg gcgcgggtgg tgtgcttgtc cttgcggttg gtgaagcccg
     1501 ccaagcggcc atagtggcgg ctgtcggcgc tggccgggtc ggcgtcgtac tcgctggcca
     1561 gcgtccgggc aatctgcccc cgaagttcac cgcctgcggc gtcggccacc ttgacccatg
     1621 cctgatagtt cttcgggctg gtttccacta ccagggcagg ctcccggccc tcggctttca
     1681 tgtcatccag gtcaaactcg ctgaggtcgt ccaccagcac cagaccatgc cgctcctgct
     1741 cggcgggcct gatatacacg tcattgccct gggcattcat ccgcttgagc catggcgtgt
     1801 tctggagcac ttcggcggct gaccattccc ggttcatcat ctggccggtg gtggcgtccc
     1861 tgacgccgat atcgaagcgc tcacagccca tggccttgag ctgtcggcct atggcctgca
     1921 aagtcctgtc gttcttcatc gggccaccaa gcgattccca cacattatac gagccggaag
     1981 cataaagtgt agctagatcc gaaggatgag ccgggctgaa tgatcgaccg agacaggccc
     2041 tgcggggctg cacacgcgcc cccacccttc gggtaggggg aaaggccgct aaagcggcta
     2101 aaagcgctcc agcgtatttc tgcggggttt ggtgtggggt ttagcgggct ttgcccgcct
     2161 ttccccctgc cgcgcagcgg tggggcggtg tgtagcctag cgcagcgaat agaccagcta
     2221 tccggcctct ggccgggcat attgggcaag ggcagcagcg ccccacaagg gcgctgataa
     2281 ccgcgcctag tggattattc ttagataatc atggatggat ttttccaaca ccccgccagc
     2341 ccccgcccct gctgggtttg caggtttggg ggcgtgacag ttattgcagg ggttcgtgac
     2401 agttattgca ggggggcgtg acagttattg caggggttcg tgacagttag ggcgcgccca
     2461 gctgtctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     2521 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ttaattaaag
     2581 cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt cgcggccgct
     2641 tctagagagc tgttgacaat taatcatccg gctcgtataa tgtgtggatc cccgtcagtg
     2701 acggagattc acacatacta gagattaaag aggagaaata ctagatgcat catcaccacc
     2761 accatggttc cggtaagatt attgcgtatg gagcgcgggt tgatgagatc caatacttta
     2821 agcagtgggc taaagacacc gggaatacct tggagtatca caccgaattt cttgatgaaa
     2881 ataccgttga atgggccaag gggttcgacg gtattaatag tttgcaaaca accccctacg
     2941 cggccggcgt gtttgaaaaa atgcacgcgt atgggatcaa attcttaacc attaggaatg
     3001 tgggcaccga caatatcgac atgaccgcca tgaaacagta tggaatccgc ttaagtaacg
     3061 tccccgccta tagtcccgcc gccattgccg aatttgcatt gacagacacc ttgtatctgc
     3121 tacggaatat ggggaaagtg caagcccaat tacaagcggg ggactatgag aaagccggca
     3181 cctttatcgg aaaagaacta ggtcaacaga ccgtcggcgt gatgggtacc ggtcacatcg
     3241 gccaggtggc gatcaaattg tttaaagggt ttggtgctaa ggttatagct tacgatccgt
     3301 atcccatgaa aggcgaccat cccgattttg attatgtgtc ccttgaagac ctgtttaaac
     3361 aaagcgatgt catcgactta cacgtgcctg gaatcgaaca gaacacgcat atcatcaatg
     3421 aggccgcctt taatctgatg aaacctggtg cgattgtgat taacaccgca cggcctaact
     3481 taattgatac gcaggccatg ctcagcaact tgaaatcagg caaactcgca ggggtgggta
     3541 tagacactta cgaatatgaa actgaagatc ttttgaacct ggcgaagcac ggttccttta
     3601 aagatccttt gtgggatgaa ttattaggga tgccgaatgt ggtattgtct cctcatatcg
     3661 cttactacac cgaaactgct gtgcataata tggtgtactt tagccttcaa catctggtgg
     3721 attttctgac taaaggcgag acttctacag aagtgaccgg cccagcaaaa taatgatact
     3781 agtagcggcc gctgcaggca tgcaagcttg cggccgcgtc gtgactggga aaaccctggc
     3841 gactagtctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
     3901 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag gggtccccaa
     3961 taattacgat ttaaatttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     4021 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     4081 agccatattc agcgtgaaac gagctgtagc cgtccgcgtc tgaacagcaa catggatgcg
     4141 gatctgtatg gctataaatg ggcgcgtgat aacgtgggtc agagcggcgc gaccatttat
     4201 cgtctgtatg gcaaaccgga tgcgccggaa ctgtttctga aacatggcaa aggcagcgtg
     4261 gcgaacgatg tgaccgatga aatggtgcgt ctgaactggc tgaccgaatt tatgccgctg
     4321 ccgaccatta aacattttat tcgcaccccg gatgatgcgt ggctgctgac caccgcgatt
     4381 ccgggcaaaa ccgcgtttca ggtgctggaa gaatatccgg atagcggcga aaacattgtg
     4441 gatgcgctgg ccgtgtttct gcgtcgtctg catagcattc cggtgtgcaa ctgcccgttt
     4501 aacagcgatc gtgtgtttcg tctggcccag gcgcagagcc gtatgaacaa cggcctggtg
     4561 gatgcgagcg attttgatga tgaacgtaac ggctggccgg tggaacaggt gtggaaagaa
     4621 atgcataaac tgctgccgtt tagcccggat agcgtggtga cccacggcga ttttagcctg
     4681 gataacctga ttttcgatga aggcaaactg attggctgca ttgatgtggg ccgtgtgggc
     4741 attgcggatc gttatcagga tctggccatt ctgtggaact gcctgggcga atttagcccg
     4801 agcctgcaaa aacgtctgtt tcagaaatat ggcattgata atccggatat gaacaaactg
     4861 caatttcatc tgatgctgga tgaatttttc taataattaa ttggaccgcg gtccgcgcgt
     4921 tgtccttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     4981 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     5041 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     5101 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     5161 gctggccgta ggccggcctc agcctgccgc cttgggccgg gtgatgtcgt acttgcccgc
     5221 cgcgaactcg gttaccgtcc agcccagcgc gaccagctcc ggcaacgcct cgcgcacccg
     5281 ctggcggcgc ttgcgcatgg tcgaaccact ggcctctgac ggccagacat agccgcacaa
     5341 ggtatctatg gaagccttgc cggttttgcc ggggtcgatc cagccacaca gccgctggtg
     5401 cagcaggcgg gcggtttcgc tgtccagcgc ccgcacctcg tccatgctga tgcgcacatg
     5461 ctggccgcca cccatgacgg cctgcgcgat caaggggttc agggccacgt acaggcgccc
     5521 gtccgcctcg tcgctggcgt actccgacag cagccgaaac ccctgccgct tgcggccatt
     5581 ctgggcgatg atggatacct tccaaaggcg ctcgatgcag tcctgtatgt gcttgagcgc
     5641 cccaccacta tcgacctctg ccccgatttc ctttgccagc gcccgatagc tacctttgac
     5701 caccatggca tcagcggtga cggcctccca cttgggttcc aggaacagcc ggagctgccg
     5761 tccgccttcg gtcttgggtt ccgggccaag cactaggcca ttaggcccag ccatggccac
     5821 cagcccttgc aggatgcgca gatcatcagc gcccagcggc tccgggccgc tgaactcgat
     5881 ccgcttgccg tcgccgtagt catacgtcac gtccagcttg ctgcgcttgc gctcgccccg
     5941 cttgagggca cggaacaggc cgggggccag acagtgcgcc gggtcgtgcc ggacgtggct
     6001 gaggctgtgc ttgttcttag gcttcaccac ggggcacccc cttgctcttg cgctgcctct
     6061 ccagcacggc gggcttgagc accccgccgt catgccgcct gaaccaccga tcagcgaacg
     6121 gtgcgccata gttggccttg ctcacaccga agcggacgaa gaaccggcgc tggtcgtcgt
     6181 ccacacccca ttcctcggcc tcggcgctgg tcatgctcga caggtaggac tgccagcgga
     6241 tgttatcgac cagtaccgag ctgccccggc tggcctgctg ctggtcgcct gcgcccatca
     6301 tggccgcgcc cttgctggca tggtgcagga acacgataga gcacccggta tcggcggcga
     6361 tggcctccat gcgaccgatg acctgggcca tggggccgc
//
