LOCUS       sequence-64076-e        4862 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 4862)
  TITLE     McLaughlin Lab KillerRed plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4862)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCS2-NXE+mem-KillerRed
FEATURES             Location/Qualifiers
     source          1..4862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    910..1044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(934..953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     988..1007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1197..1213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1197..1213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1210..1232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1221..1237
                     /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)."
     promoter        complement(join(1245..1251,1252..1269,1270..1275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1290..1311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1428..1445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1599..2187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1679..1698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2358..3149,3150..3218))
                     /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     2981..3000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3219..3323)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3349..3804)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3486..3507)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3698..3717
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3878..4862
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccggt cgccaccatg ctgtgctgta tgagaagaac
      121 caaacaggtt gaaaagaatg atgaggacca aaagatctcc gagggcggcc ccgccctgtt
      181 ccagagcgac atgaccttca aaatcttcat cgacggcgag gtgaacggcc agaagttcac
      241 catcgtggcc gacggcagca gcaagttccc ccacggcgac ttcaacgtgc acgccgtgtg
      301 cgagaccggc aagctgccca tgagctggaa gcccatctgc cacctgatcc agtacggcga
      361 gcccttcttc gcccgctacc ccgacggcat cagccatttc gcccaggagt gcttccccga
      421 gggcctgagc atcgaccgca ccgtgcgctt cgagaacgac ggcaccatga ccagccacca
      481 cacctacgag ctggacgaca cctgcgtggt gagccgcatc accgtgaact gcgacggctt
      541 ccagcccgac ggccccatca tgcgcgacca gctggtggac atcctgccca acgagaccca
      601 catgttcccc cacggcccca acgccgtgcg ccagctggcc ttcatcggct tcaccaccgc
      661 cgacggcggc ctgatgatgg gccacttcga cagcaagatg accttcaacg gcagccgcgc
      721 catcgagatc cccggcccac acttcgtgac catcatcacc aagcagatga gggacaccag
      781 cgacaagcgc gaccacgtgt gccagcgcga ggtggcctac gcccacagcg tgccccgcat
      841 caccagcgcc atcggtagcg acgaggatta aagcggccgc tctagaacta tagtgagtcg
      901 tattacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
      961 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1021 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1081 cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg
     1141 gtacccagct tttgttcccc ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1201 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1261 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1321 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1381 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2401 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2461 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2521 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2581 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2641 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2701 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2761 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2821 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2881 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2941 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3001 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3061 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3121 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3181 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3241 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3301 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     3361 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3421 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3481 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3541 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3601 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3661 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3721 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3781 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     3841 ggcgatcggt gcgggcctct tcgctattac gccagtcgac catagccaat tcaatatggc
     3901 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
     3961 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
     4021 ggacttggca ccatgccaat tcaatatggc ggacttggca ctgtgccaac tggggagggg
     4081 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata
     4141 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat
     4201 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta
     4261 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct
     4321 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc
     4381 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc
     4441 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct
     4501 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc
     4561 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg
     4621 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aaggacgcca
     4681 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
     4741 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
     4801 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
     4861 ac
//
