LOCUS       sequence-102513-        6540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MXS_chaining vector with 2 cassettes .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6540)
  AUTHORS   Sladitschek HL, Neveu PA
  TITLE     MXS-Chaining: A Highly Efficient Cloning Platform for Imaging and 
            Flow Cytometry Approaches in Mammalian Systems.
  JOURNAL   PLoS One. 2015 Apr 24;10(4):e0124958. doi: 
            10.1371/journal.pone.0124958. eCollection 2015.
  PUBMED    25909630
REFERENCE   2  (bases 1 to 6540)
  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: "PLoS One.";
            date: "2015-04-24"; volume: "10(4):e0124958. doi"; pages: " 
            10.1371/journal.pone.0124958. eCollection 2015"
COMMENT     Sequence Label: rtTA and TagBFP cassettes
FEATURES             Location/Qualifiers
     source          1..6540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             651..671
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2775..2999
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        3065..3368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3369..3572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(3634..4251,4252..4257,4258..4293,4294..4296,4297..4332,
                     4333..4335,4336..4371,4372..4380)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    4401..4625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4648..4752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4753..4821,4822..5613)
                     /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(4971..4990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5784..6372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6273..6292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgatatca aacgcgtaac tcgagtagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag ggatctcagc tggtcgaggc caccatggct cctaagaaga
      661 agaggaaggt ggtcgagagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
      721 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
      781 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
      841 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
      901 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
      961 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     1021 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     1081 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     1141 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     1201 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     1261 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     1321 tggcagtggc cagatacgcg ggcctgggcg gcggcctgaa cgtcgagagc gagctgatta
     1381 aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt
     1441 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
     1501 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca
     1561 gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg
     1621 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     1681 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     1741 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga
     1801 cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg
     1861 gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga
     1921 acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca
     1981 acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatacgcg ggcctgggcg
     2041 gcggcctgaa cgtcgaggtg agctctatct ttggggtcga gagcgagctg attaaggaga
     2101 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
     2161 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     2221 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
     2281 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
     2341 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     2401 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
     2461 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt
     2521 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
     2581 gccatctgat cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca
     2641 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
     2701 agacctacgt cgagcagcac gaggtggcag tggccagata cgcgggcctg ggcggcggcc
     2761 tgaactaagt cgagctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2821 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2881 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2941 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggt
     3001 cgagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3061 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     3541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagg
     3601 gatctcagct ggtcgaggca ggtcgaattc accatgtcta gactggacaa gagcaaagtc
     3661 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa
     3721 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     3781 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     3841 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct
     3901 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     3961 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     4021 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     4081 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     4141 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     4201 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc
     4261 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     4321 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     4381 ctaagtaagg atctgtcgag ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4441 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4501 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4561 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4621 tatggtcgac caattcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4741 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4801 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4861 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4921 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4981 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5041 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5101 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5161 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5221 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5281 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5341 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5401 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5461 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5521 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5581 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5641 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5701 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6421 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6481 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
//
