LOCUS       sequence-230588-        6921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in S. rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6921)
  TITLE     King Lab plasmids
REFERENCE   2  (bases 1 to 6921)
  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: NK691
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2213..2275
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   6224..6258
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6277..6296)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6278..6296)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6306..6323)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6306..6322)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6315..6337)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6464..6919
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6551..6570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6761..6782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtactact agccctagaa
     2221 agataatcat attgtgacgt acgttaaaga taatcatgcg taaaattgac gcatgtgttt
     2281 tatcggtctg tatatcgagg tttatttatt aatttgaata gatattaagt tttattatat
     2341 ttacacttac atactaataa taaattcaac aaacaattta tttatgttta tttatttatt
     2401 aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac aaaactttta gctagtggta
     2461 caagatctaa gcttgtaacg actgcagtct tgttgtcgtt tgtcgtcgtc gttctctgtc
     2521 tcccacccaa cttttgccaa ttgcaacagt gacttctgta caatttgcaa cagataccaa
     2581 cgtgtcgagt cagcacaaat ggcgaggaag gttcgcgtgc attgcttgcg cgcctctttg
     2641 tcatgggcgc acgctgattt gtgtgtgatt tgcaccaaat ttcctgccga cacaacttgg
     2701 gcccgtacca cacgacccgt ttctggaccc gtttctggcc cagaaacgcg aatgttgcga
     2761 aagatgcgac cacgtgctcg ctgtcgacaa cacaaactcc cgcacaatcc ccatgaaaaa
     2821 aaaagtgctt gctgggcttg tgtctggcct gtgtgcgtgg cgtggccgcg tgtggcttcg
     2881 ttcttgcttc tgctcactca gcctgttaca cgcagagtgg ctgcgagaaa caaggagagg
     2941 ggtttgcggc attggcgcat gtgcatctgc ctttttcatt gagtcagtct gcagctcaag
     3001 aggttggttt cgtcgacgga tatgagtgag cttctatgga tgatgagagg tggtggtggt
     3061 tgtatggttg gtggttggtg gttggtgcaa ggcgagccac gcgaagagcg acaaggatga
     3121 gggagaggag aggagactag gaaagaggtg aagggaaggg aagggcgagc aatgacgacg
     3181 agataaaaga cagccaccac cgcacccgca caaaagcatt gctcgcacgc atggtcgtgt
     3241 gttatgtttc acaccaccaa cataaccaat acaagccatc accaatacaa gccagcgctc
     3301 aagcacacac gcacgcacgc acacacaata cacacgcacg tgaaacgttc acattgtccg
     3361 ctcacaaatg cctttccctt ctcgtcgctt tccttgcctt tttttcttcc cctcctgact
     3421 tccttcctgc cttccttcct tcctcccctt ctccgtccac caacaccatt tatttctctg
     3481 ctctttttcc tcgtgccacg tgagcagtct tttcgcgtcg tcttcttctt tgaccccgtt
     3541 taccgtcaac accatcaaca accagcaaca tgttgagggt acgaggaaga tcgctggagg
     3601 cccggtttgt gtacatcatc gtgctggctg tggtgtacac ggccacgtac atgtacagcg
     3661 ccacatccgg tgacccaagc gtaccggttg gcggcgcaga catgccgccg tggctcgcac
     3721 agctgggacg accacgcgca gttgcagagg agctggcaga acagcaacag ctgcttgacg
     3781 cgacgcggcg cagcgacgac ggagccaaca gacacagcaa cgatggcgcc gccgcagtct
     3841 cacaacacca gcagcaccag cagcaggcgg agcacgatct ggccagcaca cggagtccaa
     3901 agacaccgac gcggcaagag gacaaaggcg tggacgagaa ggagaaaggc atgaagttcg
     3961 aggtggaggg gggcgaagat ggattcgtgc ccagggagca catgaagaat gcgatgggta
     4021 tgccagacac tgcagaggcc gacccgtgga agcggcgcag gcaggacaag tggcgcctca
     4081 tcatcttgtt ccagcacaac ttccccatcc tgaagcaggc ggtagaaggg ttccagcgtg
     4141 ccagcgacat catggtgccc agcatgatca tcgtcgacaa ctccaaggac agggatgcct
     4201 cgaacagcgt gtggcttgtg gagcgggtgg cagaggtggt cgtgccagac cgccagctaa
     4261 acttcccaga gctgcacaac tacatggcca cgctggcact cgagcgccgg ctcgaattct
     4321 acttctgggc ccacgcagac aactacgtgc tgccgctgga gcctggccgc gacatgggca
     4381 aggacgccat cgactgcctg cgcgagcaga tggcgcggtt ccccaactgg ggcatgatcc
     4441 tcttcgccta cgaccacctg gccgcctacc gcacgcagac gccagtccag gtgccatggg
     4501 acccacacgt gttccagtac ggctcggagt gcgatgcgta tgggcgcatc cgcgatgcgg
     4561 gctacgacgc gcgcgcgtgc aagattcact actcgtacga catgaagcgg gtgattggca
     4621 tcctggacac ggactcgtac gaccgggtca aggcgaagct ggacgaggag gcaaaggaca
     4681 agtccaagcg caaccagtgg cgcgagggag ccatgtccga gaaggagcag caatggcgca
     4741 agcgcatgaa ggaggcgtca cgcgcatact tggccgagaa gtggggcgag gtgaagtgca
     4801 agctgcgcgg tgtgccgtgc cacaagccct ggccttactg cccaacgtgc cccacagacc
     4861 ttccccagtg ctacggcaag gaggcgacgt gggagcaact cgaccacatt cacgccaggg
     4921 tgcaccaggt gtttgacaat gatcccgaca agccaccacc gctcgaggca ggcggtagcg
     4981 gtggctccat ggtctccaag ggcgaggaga ccacgatggg cgtcatcaag cccgacatga
     5041 agatcaagct caagatggag ggcaacgtca acggccacgc cttcgtcatc gagggcgagg
     5101 gcgagggcaa gccctacgac ggcaccaaca ccatcaacct tgaggtcaaa gaaggtgctc
     5161 ctctgccctt tagctacgac atcctcacca ccgccttctc ctacggcaac cgcgccttca
     5221 ccaagtaccc cgacgacatc cccaactact tcaagcagtc cttccccgag ggctactcct
     5281 gggagcgcac catgaccttc gaggacaagg gcatcgtcaa ggtcaagtcc gacatctcta
     5341 tggaggagga ctccttcatc tacgagatcc acctcaaggg cgagaacttc cctcccaacg
     5401 gccccgtcat gcagaaggag actacgggtt gggacgcctc caccgagcgc atgtacgtcc
     5461 gcgacggtgt tctcaagggc gacgtgaaga tgaaacttct ccttgagggc ggcggccacc
     5521 accgcgtcga cttcaagacc atctaccgcg ccaagaaggc cgtcaagctc cccgactacc
     5581 acttcgtcga ccaccgcatc gagatcctca accacgacaa ggactacaac aaagtcactg
     5641 tctacgagat cgccgtcgcc cgcaactcta ccgacggcat ggacgagctt tacaaggctt
     5701 aagcgcctcg ttggtcattg ccaccttgtt gcctcttttg ctgttgtcct cgtgtcgacg
     5761 cgtgccacgt gcgcttgttg tgagttgatt ctgcttcttg tgcacgtcat gccgagccct
     5821 tgggccccgc aataaaccca ctatgccagt gtaactatcc ttccccccaa tgtataactg
     5881 tgacgatcaa tgagcggtgc ccgagatctc ctgcaggggt accgctagct ctagcactag
     5941 tggccggccg atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa
     6001 catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat
     6061 catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt
     6121 gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac
     6181 ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg
     6241 cagactatct ttctagggct agatagctcc aattcgccct atagtgagtc gtattacgcg
     6301 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     6361 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     6421 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     6481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6661 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6721 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6781 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6841 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6901 atattaacgc ttacaattta g
//
