LOCUS       sequence-310569-        7108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive transcription of sacA crRNA and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  TITLE     Cui Lab Plasmids
REFERENCE   2  (bases 1 to 7108)
  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"
COMMENT     Sequence Label: pADsacA
FEATURES             Location/Qualifiers
     source          1..7108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..767
                     /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
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(84..112)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     707..727
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(921..940)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1207..1225
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4712..4734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4820..4960
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4875..4894
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4975..4992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5226..5245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5905..6696,6697..6765))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6528..6547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6766..6870)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7028..7099
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agatttaaga aggagatata catatgagta
       61 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      121 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
      181 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
      241 ctttcggtta tggtgttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact
      301 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      361 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
      421 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
      481 acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag
      541 ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
      601 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca
      661 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
      721 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa atgtccagac
      781 ctgcaggcat gcaagcttga gtaggacaaa tccgccgagc ttcgacgaga ttttcaggag
      841 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
      901 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
      961 ccgttcagaa caaagaatac aagaaaatat ttacaaaaaa tcaatttaac aattccttaa
     1021 aacatgcagg aattgacgat ttaaacaata ttagctttga acaattctta tctcttttca
     1081 atagctataa attatttaat aagtaagtta agggatgcat aaactgcatc ccttaacttg
     1141 tttttcgtgt gcctattttt tgtgaatcgc taagaaacca ttattatcat gacattaacc
     1201 tataaaaata ggcgtatcac gaggcccttt cgtctacata agtgtccaaa ttccgacatt
     1261 catttcagat ttcccggtcg cactggctgt tacttcatga tttcccggga aaggaaaata
     1321 tcttgcacta aagacctctt ctccgttatt gataaagatt tccacagatg acgcgtcgag
     1381 aaacatgtgt acggtatgca aatccttgat gcggcaatgt ctcgcttctg ttctttttcc
     1441 gtcaaagctc tttcgttcca gcgtaacaat cccctcgtct ttatggaagg aaaacgtagc
     1501 cgttccgcgg atactgattg aaaagccaga ctccgtatga atgtcttcca gtagaatctc
     1561 agttctttca ggtttttcca ctggaagcgc accagatgat ccatgcatgt tgatgtgtat
     1621 ttttttctca tttctgcgca tggcttttag ctcaggcagc ggctgctgaa tcagcttctg
     1681 tcctgaaagc gtcagctgtc tcggcagcgt catgcagtga atccagtggc agtcaatggt
     1741 cggatgggac ccttcatcct gatcaggcac cgccatccat gcaaataaaa tccgccttcc
     1801 ctgatcgtct tcaagtgttt gcggcgcgta aaaaaatctc cactctgtca ggttatcaga
     1861 agcaaacaac acagcctgcc ctttcaaatt ctctgtttgc gcaccaatca ccatatacca
     1921 tgtgccttca tgctcccata ccttcggatc gcgaaaatgc gccgtatatc cttccggaag
     1981 gcgggcgacg acaccctttt tctcaaagga cagcccgtcg tcagaaacag caaggcattg
     2041 atacgtttcc cgatttccat cctgatccct gacatttcct gtgtaaaaaa gatagagccg
     2101 atcgtctttc gtgacagcgc tgcccgagta gcagccgttt ttatcaaacc aatcactcgg
     2161 agccagcgca atctcttccc gcttccaatt cacaacatcc tgtgtcgtat aatgccccca
     2221 aaattttgcg ccgtgccccg tctgaaacgg ctgccactga aagaatacat gatagctgcc
     2281 cttccaataa atcacgccat tcgggtcatt cagcagccca accggcggca tgatatgaaa
     2341 atgctggcgg tgcggatcgc tgttagcgat gggctctttt ttctccactt cttcataagc
     2401 ccggcgacga agctcctggt catgtgctgt catcaaaaca tatttcaaca caatacaaat
     2461 gggttagtta aaaaagcagg ccttctaaag gtctgctttt tttatttgat tatgtaattt
     2521 ttaatgccag gatgccaata agccataacc tcaaatgcac catttgcaac ctcgtcatct
     2581 tcttcctcaa tcttgaccag atcgccgtcc cccgcatcac caagattcag ctctttatgt
     2641 atctccttca aaatgccacc gtatccaatt aaccttcgag ctgccaacgc atcatccaag
     2701 taaagcaccg tgttcagatt gtcttcagtc accttattac cgcgcacaac atccgtatcc
     2761 ttaaccggat atttagagat ttcgagaaca gctttttgct ccatcattgc gttccgcaca
     2821 tcgttttcaa tctgttcagc gtcaatctta gctttacctt tcactcgacg aatatcgaca
     2881 attggagtgt aatccaattt catcgccttt ttccaaaggc tcgtccactc cgcctgctta
     2941 atatagtttt tcccaaaata atttttcctt actggtatca acacatgaaa atgaggatga
     3001 tatgtatctt cttcatgatt tttggtaatc tctaaagctc tgaaaaatcc aagaaccgaa
     3061 gtttttactt ttttgtactg gaacagtttc ctaaagcctt ccatcatcgc agaaatttgt
     3121 ggcttcagcc gttctccctt tacatttcga atcgtcagcg tgagaaaaat ccatccgcag
     3181 ccgtactgtc tattggcttc ctctacgatc aacttattgt gataagcaat ttttaacgac
     3241 ctgcgccacg cacacatcgg acataacctc actttacaaa aatgggcttg atacagtttt
     3301 aacttgcccg tctccgggtc tctcttaaac gaaagatact ctgcacaact aattagtttt
     3361 tcagcctttt tgccatagta aggtgcccca atcttactct ctaacgcttc gtaatgctcc
     3421 gccatgaggt tcgtccgtct ctttttcccc ttccaatccc gctttttacc tgttgcggtt
     3481 ttatcttcga ggatgctata atcattttca gatgaataaa tcaacaaaaa aactccttct
     3541 gagctagttc tctagcattc tattattttg attcgacacc ttaataatag cagaaggagt
     3601 ttttacctgt caaagaacca tcaaaccctt gatacacaag gctttgacct aattttgaaa
     3661 aatgatgttg tttctatata gtatcaagat aagaaagaaa aggatttttc gctacgctca
     3721 aatcctttaa aaaaacacaa aagaccacat tttttaatgt ggtcttttat tcttcaacta
     3781 aagcacccat tagttcaaca aacgaaaatt ggataaagtg ggatattttt aaaatatata
     3841 tttatgttac agtaatattg acttttaaaa aaggattgat tctaatgaag aaagcagaca
     3901 agtaagcctc ctaaattcac tttagataaa aatttaggag gcatatcaaa tgaactttaa
     3961 taaaattgat ttagacaatt ggaagagaaa agagatattt aatcattatt tgaaccaaca
     4021 aacgactttt agtataacca cagaaattga tattagtgtt ttataccgaa acataaaaca
     4081 agaaggatat aaattttacc ctgcatttat tttcttagtg acaagggtga taaactcaaa
     4141 tacagctttt agaactggtt acaatagcga cggagagtta ggttattggg ataagttaga
     4201 gccactttat acaatttttg atggtgtatc taaaacattc tctggtattt ggactcctgt
     4261 aaagaatgac ttcaaagagt tttatgattt atacctttct gatgtagaga aatataatgg
     4321 ttcggggaaa ttgtttccca aaacacctat acctgaaaat gctttttctc tttctattat
     4381 tccatggact tcatttactg ggtttaactt aaatatcaat aataatagta attaccttct
     4441 acccattatt acagcaggaa aattcattaa taaaggtaat tcaatatatt taccgctatc
     4501 tttacaggta catcattctg tttgtgatgg ttatcatgca ggattgttta tgaactctat
     4561 tcaggaattg tcagataggc ctaatgactg gcttttataa tatgagataa tgccgactgt
     4621 actttttaca gtcggttttc taatgtcact aacctgcccc gttagtcgcc attcgccact
     4681 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4741 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4801 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     4861 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     4921 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     4981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 ctaagaaacc attattttat taacgttgat ataatttaaa ttttatttga caaaaatggg
     6961 ctcgtgttgt acaataaatg taatttctac tgttgtagat tcggccgcct cgattataac
     7021 aagatttcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     7081 tttgtcggtg aacgctctcc gtcttcaa
//
