LOCUS       sequence-405453-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An expression vector for Cre recombinase (codon usage for C. 
            reinhardtii).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4948)
  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 4948)
  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: pChlamy/Cre
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      588..1176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1077..1096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1464..1485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1500..1505,1506..1523,1524..1530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1538..1554
                     /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)."
     primer_bind     1543..1565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1562..1578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1562..1578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1599..1617
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1654..1670
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1654..1670
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2361..2381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3735..3754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3736..3754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3761..3778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3761..3777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3770..3792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3918..4373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4055..4076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4267..4286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4400..4504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4505..4573,4574..4948,1..417)
                     /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     complement(4723..4742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
       61 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      121 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      181 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      241 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
      301 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      361 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
      421 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      721 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      961 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1261 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1321 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     1381 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1441 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1501 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1561 acaggaaaca gctatgacca tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa
     1621 caaaagctgg agctccaccg cggtggcggc cgctctagaa ctagtggatc ttcgctgagg
     1681 cttgacatga ttggtgcgta tgtttgtatg aagctacagg actgatttgg cgggctatga
     1741 gggcgcggga agctctggaa gggccgcgat ggggcgcgcg gcgtccagaa ggcgccatac
     1801 ggcccgctgg cggcacccat ccggtataaa agcccgcgac cccgaacggt gacctccact
     1861 ttcagcgaca aacgagcact tatacatacg cgactattct gccgctatac ataaccactc
     1921 agctagctta agatcccatc aagcttgcat gccgggcgcg ccagaaggag cgcagccaaa
     1981 ccaggatgat gtttgatggg gtatttgagc acttgcaacc cttatccgga agccccctgg
     2041 cccacaaagg ctaggcgcca atgcaagcag ttcgcatgca gcccctggag cggtgccctc
     2101 ctgataaacc ggccaggggg cctatgttct ttactttttt acaagagaag tcactcaaca
     2161 tcttaaaatg gccaggtgag tcgacgagca agcccggcgg atcaggcagc gtgcttgcag
     2221 atttgacttg caacgcccgc attgtgtcga cgaaggcttt tggctcctct gtcgctgtct
     2281 caagcagcat ctaaccctgc gtcgccgttt ccatttgcag gagattcgag gtacccatgg
     2341 atatcgcgtc ctccggtccg ccgaagaaga agcggaaggt ggaggacccc ggtggcagag
     2401 atatcatgtc gaacctcctc accgtgcacc agaaccttcc ggctctgccc gtcgacgcca
     2461 cgagcgacga ggtccgtaag aacctgatgg acatgttccg cgaccgccag gcgttcagcg
     2521 agcacacctg gaagatgctg ctgtccgtgt gccgctcatg ggccgcttgg tgcaagctga
     2581 acaaccggaa gtggtttcca gccgaacccg aggacgtgcg cgactacctg ctgtatctgc
     2641 aggcccgagg gctggcggtg aagacgatcc agcagcacct gggccagctg aacatgctgc
     2701 atcgccgcag cggcctgcct cggccctccg acagcaatgc cgtgtcgttg gtgatgcgcc
     2761 gcattcgcaa ggagaacgtg gatgccggcg agagggcgaa gcaagcacta gcgttcgagc
     2821 gtaccgactt cgaccaggtg cgcagcctga tggagaactc ggaccgctgc caggacatcc
     2881 gcaatctggc cttcctgggc atcgcctaca acacgttgct gcgcatcgcc gagatcgccc
     2941 gcattagggt caaggacatc tcgcgcactg acggcggccg catgctgatc cacataggcc
     3001 ggaccaagac gctggtgagc actgcaggcg tcgagaaggc ccttagcctg ggcgtgacga
     3061 agctcgtgga gcgctggatc tccgtgtccg gcgttgccga tgaccccaac aactacctct
     3121 tctgccgcgt acgcaagaac ggggtggctg cgcccagtgc gaccagccag ctgtctacac
     3181 gcgccctgga gggcatcttc gaggcgaccc accggctgat ctacggggcg aaagacgaca
     3241 gcggccagcg ttacctcgcg tggtccggac actctgcacg ggttggtgct gcgcgcgata
     3301 tggctcgcgc gggcgtgtcg atcccggaga tcatgcaggc cggcggctgg accaacgtga
     3361 acattgtgat gaactacatc cggaacctgg actcggagac cggcgccatg gtccgcctgc
     3421 tggaggacgg cgactaaaga tctctgcagc ggccgccata tgattccgct ccgtgtaaat
     3481 ggaggcgctc gttgatctga gccttgcccc ctgacgaacg gcggtggatg gaagatactg
     3541 ctctcaagtg ctgaagcggt agcttagctc cccgtttcgt gctgatcagt ctttttcaac
     3601 acgtaaaaag cggaggagtt ttgcaatttt gttggttgta acgatcctcc gttgattttg
     3661 gcctctttct ccatgggcgg gctgggcgta tttgaagctt aattaactcg agggggggcc
     3721 cggtacccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     3781 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3841 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3901 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3961 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4021 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4081 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4141 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4201 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4261 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4321 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4381 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4441 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4501 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4561 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4621 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4681 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4741 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4801 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4861 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4921 gatcggagga ccgaaggagc taaccgct
//
