LOCUS       sequence-258978-        4183 bp    DNA     circular SYN 15-APR-2025
DEFINITION  cytomplasmic marker without terminator (less integration).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4183)
  TITLE     Plasmids used for tagging different cellular components.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4183)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CAMP no 3'UTR
FEATURES             Location/Qualifiers
     source          1..4183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(32..37,38..55,56..62)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    70..86
                     /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     75..97
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     94..110
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     94..110
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             700..1407
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGAHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(845..863)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1113..1132
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1307..1330
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1417..2016
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1417..1436)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1913..1933
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2124..2143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2272..2340,2341..3132)
                     /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(2490..2509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3303..3891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3792..3811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4045..4062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(4179..4183,1..17)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
       61 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      121 ccaagcttga aatgctgggc gccttgaaac tgaacctgtt cggatccggg ttgcgcgctt
      181 ccattgtttt ggtttccgtg tctgcaaggc gggtctggac gtgacgcttt gggggagagg
      241 gagagcagag atacaattct agccacgaat ttcagccaat agtcacagag gggagttgcg
      301 atcctgcgaa ttggcttagg tctgtgaggt ctttcacggt ttcgaaagca gcgcagaggg
      361 gagaccgact accaaaagcc ataacactcc accgagatat ccgcactctt cgctagatga
      421 gcttgcttct cctcttgtca caaatcagcc caatgatagt cactgttcaa caacggtgca
      481 gtgtaaatat tcgttgtgcg tccttgatga gcagaaggta cgagaagcag gaaatcattt
      541 cataatggaa aaccattcac tgtctttctg tattgcgaaa agagtgatca taggtaatcg
      601 caacggcggg tctttagaac agtcacatac cagatcatgg gcatggagac agtagagcaa
      661 ctacgcaaga caactattgt ccccgggact agtgatatca tggtgagcaa gggcgaggag
      721 gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      781 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      841 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
      901 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      961 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1021 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1081 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     1141 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     1201 cagaggctga agctgaagga cggcgcccac tacgacgccg aggtcaagac cacctacaag
     1261 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagct ggacatcacc
     1321 tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg ccgccactcc
     1381 accggcggca tggacgagct gtacaaggcg gccgccatga ccgagtacaa gcccacggtg
     1441 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     1501 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     1561 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     1621 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     1681 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     1741 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     1801 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     1861 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     1921 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     1981 cgcacctggt gcatgacccg caagcccggt gcctgacctc gactagaggg cctgcttgcc
     2041 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2101 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     2161 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     2221 cgccttctat cgccttcttg acgagttctt ctgaattgaa aaaggaagag tatgagtatt
     2281 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2341 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2401 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2461 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2521 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2581 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2641 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2701 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2761 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2821 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2881 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2941 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3001 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3061 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3421 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3661 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3721 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3781 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3841 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3901 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3961 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4021 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4081 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4141 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atg
//
