LOCUS       sequence-354727-        4844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Level 2 destination vector for Synechocystis sp. PCC 
            6803.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4844)
  AUTHORS   Taylor GM, Hitchcock A, Heap JT
  TITLE     Combinatorial assembly platform enabling engineering of genetically 
            stable metabolic pathways in cyanobacteria.
  JOURNAL   Nucleic Acids Res. 2021 Dec 2;49(21):e123. doi: 10.1093/nar/gkab791.
  PUBMED    34554258
REFERENCE   2  (bases 1 to 4844)
  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: "Nucleic 
            Acids Res."; date: "2021-12-2"; volume: "49(21):e123. doi"; pages: "
            10.1093/nar/gkab791"
COMMENT     Sequence Label: pGT270
FEATURES             Location/Qualifiers
     source          1..4844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(111..130)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             122..937
                     /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             122..937
                     /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     complement(195..214)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2309..2854
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2755..2774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2971..2988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4378..4399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4414..4419,4420..4437,4438..4444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4452..4468
                     /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     4457..4479
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4476..4492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4476..4492
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4488..4811
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    4505..4561
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4562..4579)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4562..4578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4571..4593)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4787..4806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccaggatc caaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa
       61 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
      121 tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga
      181 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
      241 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
      301 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
      361 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
      421 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
      481 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
      541 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
      601 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
      661 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
      721 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
      781 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
      841 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
      901 attgcagttt catttgatgc tcgatgagtt tttctaatct tgggggaaat taagaccaaa
      961 ctcgatgacc tccaaaaaga tgtaacttct cttaagatcg atatggcaac ggtgaaaacc
     1021 gagttatctg cggtcaggat ggagataggt acagtcaagg atgatgttaa agatgtcaaa
     1081 gggcgggcta atgctcaaat ttgggcgttg attcttgccg tcatcggagc cataattacc
     1141 accttggtgc gttttggcat tttccctaat ccctaacaaa aaagcgacca ggcttttctt
     1201 tcaattgccc gatcgccttt gatattttcc caaaggataa aagctagtcc attcagaatc
     1261 gagccttaaa gtactcccat attggctagc cccagaatta ctccagcgcc gaggatgtgg
     1321 ccaaagctag cggtgcccag cacagcccct aaaccaaagc cgccaaagaa gttagaggaa
     1381 ggcatggggg tgcccacatt ttgttgtttg atggtcaatt taccaaaggc gatcgccaaa
     1441 atgttgcaag caatcatcac cccagcaact ttagggctcc aggacagggt ggcgggaacg
     1501 gcggtggcca acaaaaagct atgcattgag attctccaga ataaagacgg tttttaaagg
     1561 gatagcccca cgctaatggg ggtctttaaa aatctcatct tacggggacg ctctgcccct
     1621 gggaaaccac cgttgcaata cttaacaaat tttcgttttt agcttggcaa atgtctttgg
     1681 caaaattggt tgatctggct taaatcgtca gttatttgcc ctggaatagt ctggggacgg
     1741 gcaattctga tcagatttac ccccaacgct tccgccactt tttgcttaac caattctccc
     1801 ccctgggcac cggaggcttt agttaccacc ccttgaattt gccattgttg ccacagggct
     1861 ttttccaatg gttcggctac gggggggcgc aaagcaatga tacggtcgga agtaaaccca
     1921 gcggcgatcg cctgggctag ggcttgggga tagggcagaa tacgggcaaa tagaacaaat
     1981 ctggagccgg tgagcgtggg actcgcggta tcattgcagc actggggcca gatggtaagc
     2041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2281 cgtcagaccc cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     2341 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     2401 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     2461 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     2521 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     2581 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     2641 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     2701 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     2761 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     2821 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     2881 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     2941 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     3001 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     3061 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg
     3121 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc
     3181 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     3241 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcggtgg
     3301 gtcaatccct cggtttggcg atcgccttgg tgtttgtcat tgctggtatt gcctttaaaa
     3361 tttccgctgt acccttccac caatggaccc ccgacgttta tgaaggttct cccaccccgg
     3421 tggtggcttt tctttcggtg ggctctaagg cggccggttt tgccgtagct atccgattac
     3481 tagtgactgc ctttggtggt attaccgatg agtggcacgt tattttcacc gctctggccg
     3541 tgttgagcat ggtgctgggc aacgtggtgg ctttagccca aaccagcatg aaacggatgt
     3601 tggcctactc ttccatcggt caagcaggct ttgtgatgat tggcctagtg gccggcagtg
     3661 aagatggtta cgccagcatg gttttctaca tgctcatcta tctgtttatg aacctggggg
     3721 cgtttagttg cattattctc ttcaccctcc gcactggcag tgaccaaatt agtgattacg
     3781 ctggtctgta ccacaaagac cccttgttaa ccttgggctt gagcatttgt ttattatcct
     3841 tggggggcat tcctcctctg gcgggctttt tcggcaaaat ttacatcttc tgggccggtt
     3901 ggcaatcggg attgtatggc ctagtcctac ttggtctggt taccagtgta gtttccatct
     3961 actactacat ccgggtggtg aaaatgatgg tggtgaagga gccccaggaa atgtccgaag
     4021 taatcaaaaa ttacccggcc atcaaatgga atttacccgg catgcgtccc ctacaggtgg
     4081 gcattgtcgc tactttggtt gctacctcgc tggcaggtat tctggctaat cccctcttta
     4141 acctcgccac cgattccgtg gtcagcacca agatgttgca gacagccctc cagcaaacag
     4201 gagaaactcc ggcgatcgcc atttcccatg atttacccta ggggtatcag gaaatattgc
     4261 tttgcaggca aaagccaatg agtgtaacta tagaaaccga tttaaaggag actgcagtta
     4321 tggaagacaa tgccggagag agacccagta ccagtagggc agtgagcgca acgcaattaa
     4381 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4441 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4501 cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg agctcgaatt
     4561 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4621 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4681 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     4741 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     4801 atgccgcata gtttcctttc cggtctcata ctactaaagt cttc
//
