LOCUS       sequence-405460-        6931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Gaussia luciferase (gLuc) and spectinomycin resistance
            (Spec) genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6931)
  AUTHORS   Huang G, Kawabe Y, Shirakawa K, Akiyama T, Kamihira M
  TITLE     Novel transgenic Chlamydomonas reinhardtii strain with retargetable 
            genomic transgene integration using Cre-loxP system.
  JOURNAL   J Biosci Bioeng. 2021 Nov;132(5):469-478. doi: 
            10.1016/j.jbiosc.2021.07.006. Epub 2021 Aug 19.
  PUBMED    34420898
REFERENCE   2  (bases 1 to 6931)
  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"; doi: 
            "10.1016/j.jbiosc.2021.07.006"; journalName: "J Biosci Bioeng"; 
            date: "2021-11"; volume: "132"; issue: "5"; pages: "469-478"
COMMENT     Sequence Label: pHR-LS
FEATURES             Location/Qualifiers
     source          1..6931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..50)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     241..260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     474..496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     488..505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     489..505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     515..534
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        515..533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1342..1353
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2190..2201
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2461..2484
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4632..4652)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4632..4650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4671..4687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4684..4706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4695..4711
                     /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)."
     promoter        complement(join(4719..4725,4726..4743,4744..4749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4764..4785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6623,6624..6692))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6693..6797)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6823..6931,1..347))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
       61 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
      361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      481 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
      541 ggagctccac cgcggtggcg gccgctctag agctgaggct tgacatgatt ggtgcgtatg
      601 tttgtatgaa gctacaggac tgatttggcg ggctatgagg gcgggggaag ctctggaagg
      661 gccgcgatgg ggcgcgcggc gtccagaagg cgccatacgg cccgctggcg gcacccatcc
      721 ggtataaaag cccgcgaccc cgaacggtga cctccacttt cagcgacaaa cgagcactta
      781 tacatacgcg actattctgc cgctatacat aaccactcag ctagcttaag atcccatcac
      841 cggtgcatgc cgggcgcgcc agaaggagcg cagccaaacc aggatgatgt ttgatggggt
      901 atttgagcac ttgcaaccct tatccggaag ccccctggcc cacaaaggct aggcgccaat
      961 gcaagcagtt cgcatgcagc ccctggagcg gtgccctcct gataaaccgg ccagggggcc
     1021 tatgttcttt acttttttac aagagaagtc actcaacatc ttaaaatggc caggtgagtc
     1081 gacgagcaag cccggcggat ccataacttc gtataaagta tcctatacga acggtaagat
     1141 ctaggcagcg tgcttgcaga tttgacttgc aacgcccgca ttgtgtcgac gaaggctttt
     1201 ggctcctctg tcgctgtctc aagcagcatc taaccctgcg tcgccgtttc catttgcagg
     1261 atgcatatgg cgcgtactgg cgctctactc ctggtcgcgc tggcgcttgc gggctgcgcg
     1321 caggcttgca gatctgacgt catcgagggc aggaagccga cggagaacaa cgaggacttc
     1381 aacattgtcg ccgtggcgtc caactttgcc accaccgatc tggacgccga ccgcggcaag
     1441 ctgccgggca agaagctgcc cctggaggtc ctcaaggaaa tggaggcgaa cgcccggaag
     1501 gccggctgca cccgcggctg cctgatctgc ctgtcccaca tcaagtgcac gccgaagatg
     1561 aagaagttca ttcccggccg gtgagcttgc ggggttgcga gcaacactcc agcaacgaac
     1621 agtgcccaag tcaggaatct gcagtcagcc tgggctttcg gcggcttttt cttgggcaaa
     1681 cagcttgcac tcatgccagc gcggcttgtc cagcctcact tgagctttcc agctgctacc
     1741 agccgggcta tacgacagcg acagagccat agcgtggaat cacttatttg ggttgccgaa
     1801 gtagcggtcg gagcgtgagt tcttggtcaa gccgcccctt atccggttcc tgtccgtgtc
     1861 tttgtccctc gttcaccctt cgcggcaccc ttcatcccct tgcttgcagg tgccacacct
     1921 acgagggcga caaggagtcc gcgcagggcg gcattggcga ggcgatcgtg gacatccccg
     1981 agattccggg cttcaaggac ctggagccga tggagcagtt catcgcccag gtggacctgt
     2041 gcgtggactg cacgaccggc tgtctgaagg gcctggccaa tgtgcagtgt tccgacctgc
     2101 tcaagaagtg gctgccccag cgctgtgcca ccttcgccag caagatccag ggccaggtgg
     2161 acaagattaa gggcgctggc ggcgatacga tcgagggcag ggatatctaa gtgagtctgg
     2221 cgagagcccg acgggtccac tgtggcactg ggttagcttt tggcacacgg gtccactgtg
     2281 gcactggtta gcttggcacc gggacagcgc ctatctcacc gcggggaact gacgcatacc
     2341 cctgctcgtg cttcagcacg gaaaagcaag gggcccaatt ccatctttgg tggttctgtg
     2401 cgctggtgac tgaacctctt ctccctccca tttcccgtgc gcccgcagcc cggggaattc
     2461 tggagccacc cgcagttcga gaagtaaccg ctccgtgtaa atggaggcgc tcgttgatct
     2521 gagccttgcc ccctgacgaa cggcggtgga tggaagatac tgctctcaag tgctgaagcg
     2581 gtagcttagc tccccgtttc gtgctgatca gtctttttca acacgtaaaa agcggaggag
     2641 ttttgcaatt ttgttggttg taacgatcct ccgttgattt tggcctcttt ctccatgggc
     2701 gggctgggcg tatttgaagc gactagtacg cgtgctgagg cttgacatga ttggtgcgta
     2761 tgtttgtatg aagctacagg actgatttgg cgggctatga gggcggggga agctctggaa
     2821 gggccgcgat ggggcgcgcg gcgtccagaa ggcgccatac ggcccgctgg cggcacccat
     2881 ccggtataaa agcccgcgac cccgaacggt gacctccact ttcagcgaca aacgagcact
     2941 tatacatacg cgactattct gccgctatac ataaccactc agctagctta agatcccatc
     3001 cctagggcat gccgggcgcg ccagaaggag cgcagccaaa ccaggatgat gtttgatggg
     3061 gtatttgagc acttgcaacc cttatccgga agccccctgg cccacaaagg ctaggcgcca
     3121 atgcaagcag ttcgcatgca gcccctggag cggtgccctc ctgataaacc ggccaggggg
     3181 cctatgttct ttactttttt acaagagaag tcactcaaca tcttaaaatg gccaggtgag
     3241 tcgacgagca agcccggcgg atccataact tcgtataaag tatcctatac gaacggtaag
     3301 atctaggcag cgtgcttgca gatttgactt gcaacgcccg cattgtgtcg acgaaggctt
     3361 ttggctcctc tgtcgctgtc tcaagcagca tctaaccctg cgtcgccgtt tccatttgca
     3421 ggaagcttat gcgcgaggcc gtgatcgccg aggtgagcac ccagctgagc gaggtggtgg
     3481 gcgtgatcga gcgccacctg gagcccaccc tgctggccgt gcacctgtac ggcagcgccg
     3541 tggacggcgg cctgaagccc cacagcgaca tcgacctgct ggtgaccgtg accgtgcgcc
     3601 tggacgagac cacccgccgc gccctgatca acgacctgct ggagaccagc gccagccccg
     3661 gcgagagcga gatcctgcgc gccgtggagg tgaccatcgt ggtgcacgac gacatcatcc
     3721 cctggcgcta ccccgccaag cgcgagctgc agttcggcga gtggcagcgc aacgacatcc
     3781 tggccggcat cttcgagccc gccaccatcg acatcgacct ggccatcctg ctgaccaagg
     3841 cccgcgagca cagcgtggcc ctggtgggcc ccgccgccga ggtgagtcga cgagcaagcc
     3901 cggcggatca ggcagcgtgc ttgcagattt gacttgcaac gcccgcattg tgtcgacgaa
     3961 ggcttttggc tcctctgtcg ctgtctcaag cagcatctaa ccctgcgtcg ccgtttccat
     4021 ttgcaggagc tgttcgaccc cgtgcccgag caggacctgt tcgaggccct gaacgagacc
     4081 ctgaccctgt ggaacagccc ccccgactgg gccggcgacg agcgcaacgt ggtgctgacc
     4141 ctgagccgca tctggtacag cgccgtgacc ggcaagatcg cccccaagga cgtggccgcc
     4201 gactgggcca tggagcgcct gcccgcccag taccagcccg tgatcctgga ggcccgccag
     4261 gcctacctgg gccaggagga ggaccgcctg gccagccgcg ccgaccagct ggaggagttc
     4321 gtgcactacg tgaagggcga gatcaccaag gtggtgggca agtaactcga gtgaccgctc
     4381 cgtgtaaatg gaggcgctcg ttgatctgag ccttgccccc tgacgaacgg cggtggatgg
     4441 aagatactgc tctcaagtgc tgaagcggta gcttagctcc ccgtttcgtg ctgatcagtc
     4501 tttttcaaca cgtaaaaagc ggaggagttt tgcaattttg ttggttgtaa cgatcctccg
     4561 ttgattttgg cctctttctc catgggcggg ctgggcgtat ttgaagcgga cccggtaccc
     4621 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     6841 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     6901 aaatcggcaa aatcccttat aaatcaaaag a
//
