LOCUS       sequence-140264-        6220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6220)
  AUTHORS   Cowley SC, Av-Gay Y
  TITLE     Monitoring promoter activity and protein localization in 
            Mycobacterium spp. using green fluorescent protein.
  JOURNAL   Gene. 2001 Feb 21;264(2):225-31.
  PUBMED    11250077
REFERENCE   2  (bases 1 to 6220)
  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: "Gene."; 
            date: "2001-02-21"; volume: "264(2)"; pages: "225-31"
COMMENT     Sequence Label: pSC300
FEATURES             Location/Qualifiers
     source          1..6220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             432..443
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             503..1216
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(533..561)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1156..1176
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1235..1251)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1236..1252)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     rep_origin      5451..6039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5940..5959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6193..6210
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacc agatctggct
      241 cgcaccgcgg tggcggccgc tctagacagc ctgggggcgt cctggtagcc tatgccgtcc
      301 gccctggggc atcgacccca aggtcgttgt tgcgacgcga gcggtcatgg agcagggttg
      361 acttgtcaag ctagagccag cccatcgcgt gggaggcacc cgcgcgaaaa gaaacatcgg
      421 acgatcattt catcgaagga aggaatgccg tgggatcccc cgggctgcag gaattcgata
      481 tcaagctgta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc
      541 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      601 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      661 acctgttcca tggccaacac ttgtcactac tttgacttat ggtgttcaat gcttttcaag
      721 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
      781 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa
      841 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
      901 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
      961 ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga
     1021 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     1081 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
     1141 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     1201 ggatgaacta tacaaatagc attcgtaagc ttatcgatac cgtcgacctc gagggggggc
     1261 ccccggggat catcgagccg agaacgttat cgaagttggt catgtgtaat cccctcgttt
     1321 gaactttgga ttaagcgtag atacaccctt ggacaagcca gttggattcg gagacaagca
     1381 aattcagcct taaaaagggc gaggccctgc ggtggtggaa caccgcaggg cctctaaccg
     1441 ctcgacgcgc tgcaccaacc agcccgcgaa cggctggcag ccagcgtaag gcgcggctca
     1501 tcgggcggcg ttcgccacga tgtcctgcac ttcgagccaa gcctcgaaca cctgctggtg
     1561 tgcacgactc acccggttgt tgacaccgcg cgcggccgtg cgggctcggt ggggcggctc
     1621 tgtcgccctt gccagcgtga gtagcgcgta cctcacctcg cccaacaggt cgcacacagc
     1681 cgattcgtac gccataaagc caggtgagcc caccagctcc gtaagttcgg gcgctgtgtg
     1741 gctcgtaccc gcgcattcag gcggcagggg gtctaacggg tctaaggcgg cgtgtacggc
     1801 cgccacagcg gctctcagcg gcccggaaac gtcctcgaaa cgacgcatgt gttcctcctg
     1861 gttggtacag gtggttgggg gtgctcggct gtcgctggtg ttccaccacc agggctcgac
     1921 gggagagcgg gggagtgtgc agttgtgggg tggcccctca gcgaaatatc tgacttggag
     1981 ctcgtgtcgg accatacacc ggtgattaat cgtggtctac taccaagcgt gagccacgtc
     2041 gccgacgaat ttgagcagct ctggctgccg tactggccgc tggcaagcga cgatctgctc
     2101 gaggggatct accgccaaag ccgcgcgtcg gccctaggcc gccggtacat cgaggcgaac
     2161 ccaacagcgc tggcaaacct gctggtcgtg gacgtagacc atccagacgc agcgctccga
     2221 gcgctcagcg cccgggggtc ccatccgctg cccaacgcga tcgtgggcaa tcgcgccaac
     2281 ggccacgcac acgcagtgtg ggcactcaac gcccctgttc cacgcaccga atacgcgcgg
     2341 cgtaagccgc tcgcatacat ggcggcgtgc gccgaaggcc ttcggcgcgc cgtcgacggc
     2401 gaccgcagtt actcaggcct catgaccaaa aaccccggcc acatcgcctg ggaaacggaa
     2461 tggctccact cagatctcta cacactcagc cacatcgagg ccgagctcgg cgcgaacatg
     2521 ccaccgccgc gctggcgtca gcagaccacg tacaaagcgg ctccgacgcc gctagggcgg
     2581 aattgcgcac tgttcgattc cgtcaggttg tgggcctatc gtcccgccct catgcggatc
     2641 tacctgccga cccggaacgt ggacggactc ggccgcgcga tctatgccga gtgccacgcg
     2701 cgaaacgccg aattcccgtg caacgacgtg tgtcccggac cgctaccgga cagcgaggtc
     2761 cgcgccatcg ccaacagcat ttggcgttgg atcacaacca agtcgcgcat ttgggcggac
     2821 gggatcgtgg tctacgaggc cacactcagt gcgcgccagt cggccatctc gcggaagggc
     2881 gcagcagcgc gcacggcggc gagcacagtt gcgcggcgcg caaagtccgc gtcagccatg
     2941 gaggcattgc tatgagcgac ggctacagcg acggctacaa ccggcagccg actgtccgca
     3001 aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac cggactatcc gaacgccacg
     3061 tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc cgagcaggct gcacgccgcg
     3121 aacgcatccg cgcctatcac gacgacgagg gccactcttg gccgcaaacg gccaaacatt
     3181 tcgggctgca tctggacacc gttaagcgac tcggctatcg ggcgaggaaa gagcgtgcgg
     3241 cagaacagga agcggctcaa aaggcccaca acgaagccga caatccaccg ctgttctaac
     3301 gcaattgggg agcgggtgtc gcgggggttc cgtggggggt tccgttgcaa cgggtcggac
     3361 aggtaaaagt cctggtagac gctagttttc tggtttgggc catgcctgtc tcgttgcgtg
     3421 tttcgttgcg cccgttttga ataccagcca gacgagacgg ggttctacga atcttggtcg
     3481 ataccaagcc atttccgctg aatatcgggg agctcaccgc cagaatcggt ggttgtggtg
     3541 atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat ccgtggccac tctcgttgca
     3601 cggttcgttg tgccgttaca ggccccgttg acagctcacc gaacgtagtt aaaacatgct
     3661 ggtcaaacta ggtttaccaa cgatacgagt cagctcatct agggccagtt ctaggcgttg
     3721 ttcgttgcgc ggttcgttgc gcatgtttcg tgtggttgct agatggctcc gcaaccacac
     3781 gcttcgaggt tgagtgcttc cagcacgggc gcgatccaga agaacttcgt cgtgcgactg
     3841 tcctcgttga tccttgccga gctgggatgg aagcccggcc gacccaccct ggaggagatg
     3901 atcgaggatg ccagggcctt tcacgcccgc cgctgctgag cgtccgccgc cgggcccgca
     3961 ccgccgtcgg ccggcccgct ccgggctcgc agcagcgggc ttcggcgcgg gcccggggct
     4021 cccgggccgc cgggcggggc tccgcccggc ggccgccggg ggccgggggc ggcgccgggc
     4081 ggcccggggc gtcaggcgcc gggggcggtg tccggcggcc cccagaggaa ctgcgccagt
     4141 tcctccggat cggtgaagcc ggagagatcc agcggggtct cctcgaacac ctcgaagtcg
     4201 tgcaggaagg tgaaggcgag cagttcgcgg gcgaagtcct cggtccgctt ccactgcgcc
     4261 ccgtcgagca gcgcggccag gatctcgcgg tcgccccgga aggcgttgag atgcagttgc
     4321 accaggctgt agcgggagtc tcccgcatag acgtcggtga agtcgacgat cccggtgacc
     4381 tcggtcgcgg ccaggtccac gaagatgttg gtcccgtgca ggtcgccgtg gacgaaccgg
     4441 ggttcgcggc cggccagcag cgtgtccacg tccggcagcc agtcctccag gcggtccagc
     4501 agccggggcg agaggtagcc ccacccgcgg tggtcctcga cggtcgccgc gcggcgttcc
     4561 cgcagcagtt ccgggaagac ctcggaatgg ggggtgagca cggtgttccc ggtcagcggc
     4621 accctgtgca gccggccgag cacccggccg agttcgcggg ccagggcgag cagcgcgttc
     4681 cggtcggtcg tgccgtccat cgcggaccgc caggtggtgc cggtcatccg gctcatcacc
     4741 aggtagggcc acggccaggc tccggtgccg ggccgcagct cgccgcggcc gaggaggcgg
     4801 ggcaccggca ccggggcgtc cgccaggacc gcgtacgcct ccgactccga cgcgaggctc
     4861 tccggaccgc accagtgctc gccgaacagc ttgatcaccg ggccgggctc gccgaccagt
     4921 acggggttgg tgctctcgcc gggcacccgc agcaccggcg gcaccggcag cccgagctcc
     4981 tccagggctc ggcgggccag cggctcccag aattcctggt cgttccgcag gctcgcgtag
     5041 gaatcatccg aatcaatacg gtcgagaagt aacagggatt cttgtgtcac agcggacctc
     5101 tattcacagg gtacgggccg gcttaattcc gcacggccgg tcgcgacacg gcctgtccgc
     5161 accgcggatc aggcgttgac gatgacgggc tggtcggcca cgtcggggag tcaggcaact
     5221 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5281 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5341 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5401 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5461 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5521 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5581 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5641 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5701 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5761 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5821 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5881 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5941 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6001 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6061 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6121 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6181 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
