LOCUS       sequence-233741-        5344 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 5344)
  TITLE     King Lab plasmids
REFERENCE   2  (bases 1 to 5344)
  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: NK676
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1825..1845
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1932..1988
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2937..2954)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2937..2953)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2946..2968)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3162..3181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3281..3303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3341..3359)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3427..3531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3532..3600,3601..4392)
                     /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(3750..3769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4563..5151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5052..5071
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5305..5322
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgta acgactgcag
      241 tcttgttgtc gtttgtcgtc gtcgttctct gtctcccacc caacttttgc caattgcaac
      301 agtgacttct gtacaatttg caacagatac caacgtgtcg agtcagcaca aatggcgagg
      361 aaggttcgcg tgcattgctt gcgcgcctct ttgtcatggg cgcacgctga tttgtgtgtg
      421 atttgcacca aatttcctgc cgacacaact tgggcccgta ccacacgacc cgtttctgga
      481 cccgtttctg gcccagaaac gcgaatgttg cgaaagatgc gaccacgtgc tcgctgtcga
      541 caacacaaac tcccgcacaa tccccatgaa aaaaaaagtg cttgctgggc ttgtgtctgg
      601 cctgtgtgcg tggcgtggcc gcgtgtggct tcgttcttgc ttctgctcac tcagcctgtt
      661 acacgcagag tggctgcgag aaacaaggag aggggtttgc ggcattggcg catgtgcatc
      721 tgcctttttc attgagtcag tctgcagctc aagaggttgg tttcgtcgac ggatatgagt
      781 gagcttctat ggatgatgag aggtggtggt ggttgtatgg ttggtggttg gtggttggtg
      841 caaggcgagc cacgcgaaga gcgacaagga tgagggagag gagaggagac taggaaagag
      901 gtgaagggaa gggaagggcg agcaatgacg acgagataaa agacagccac caccgcaccc
      961 gcacaaaagc attgctcgca cgcatggtcg tgtgttatgt ttcacaccac caacataacc
     1021 aatacaagcc atcaccaata caagccagcg ctcaagcaca cacgcacgca cgcacacaca
     1081 atacacacgc acgtgaaacg ttcacattgt ccgctcacaa atgcctttcc cttctcgtcg
     1141 ctttccttgc ctttttttct tcccctcctg acttccttcc tgccttcctt ccttcctccc
     1201 cttctccgtc caccaacacc atttatttct ctgctctttt tcctcgtgcc acgtgagcag
     1261 tcttttcgcg tcgtcttctt ctttgacccc gtttaccgtc aacaccatca acaaccagca
     1321 acatgtctgg tacggagtac aagcctaccg tccgcctcgc tacccgtgac gacgtgcccc
     1381 gtgctgtccg taccctcgcc gctgccttcg ctgactaccc tgctacccgt cacaccgtcg
     1441 accctgaccg ccacatcgag cgtgtcaccg agctgcagga gctgttcctc acccgcgtcg
     1501 gcctcgacat cgggaaggtc tgggtcgccg acgacggtgc tgctgtcgct gtctggacca
     1561 cccctgagtc tgtcgaggcc ggtgctgtct tcgctgagat cggcccccgc atggctgagc
     1621 tgtctggctc ccgtctcgcc gctcagcagc agatggaggg cctcctcgcc ccccaccgcc
     1681 ccaaggagcc cgcctggttc ctcgccaccg tcggcgtctc tcctgaccac cagggcaagg
     1741 gtctcggctc tgccgtcgtc ctccctggcg tcgaggccgc tgagcgtgct ggtgtccctg
     1801 ctttcctcga gacctccgct cctcgcaacc tccccttcta cgagcgtctc ggcttcaccg
     1861 tcaccgccga cgtggaggtc cccgagggtc ctcgtacctg gtgcatgacc cgcaagcccg
     1921 gtggcagcgg tgccaccaac ttttctctgc tcaagcaggc tggcgacgtt gaagagaacc
     1981 ccggacccat ggtctccaag ggcgaggaga ccacgatggg cgtcatcaag cccgacatga
     2041 agatcaagct caagatggag ggcaacgtca acggccacgc cttcgtcatc gagggcgagg
     2101 gcgagggcaa gccctacgac ggcaccaaca ccatcaacct cgaggtcaag gagggcgccc
     2161 ccctcccctt ctcctacgac atcctcacca ccgccttcgc ctacggcaac cgcgccttca
     2221 ccaagtaccc cgacgacatc cccaactact tcaagcagtc cttccccgag ggctactcct
     2281 gggagcgcac catgaccttc gaggacaagg gcatcgtcaa ggtcaagtcc gacatctcta
     2341 tggaggagga ctccttcatc tacgagatcc acctcaaggg cgagaacttc ccccccaacg
     2401 gccccgtcat gcagaagaag accaccggct gggacgcctc caccgagcgc atgtacgtcc
     2461 gcgacggcgt cctcaagggc gacgtgaagc acaagctcct cctcgagggc ggcggccacc
     2521 accgcgtcga cttcaagacc atctaccgcg ccaagaaggc cgtcaagctc cccgactacc
     2581 acttcgtcga ccaccgcatc gagatcctca accacaacaa ggactacaac aaggtcaccg
     2641 tctacgagtc cgccgtcgcc cgcaactcca ccgacggcat ggacgagctg tacaaggctt
     2701 aagcgcctcg ttggtcattg ccaccttgtt gcctcttttg ctgttgtcct cgtgtcgacg
     2761 cgtgccacgt gcgcttgttg tgagttgatt ctgcttcttg tgcacgtcat gccgagccct
     2821 tgggccccgc aataaaccca ctatgccagt gtaactatcc ttccccccaa tgtataactg
     2881 tgacgatcaa tgagcggtgc ccgtctagag gatccccggg taccgagctc gaattcactg
     2941 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3001 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3061 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     3121 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     3181 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3241 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3301 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3361 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3421 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4441 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4501 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4561 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4621 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4681 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4741 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4801 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4861 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4921 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4981 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5041 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5101 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5161 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat cttctttcct
     5221 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5281 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5341 atac
//
