LOCUS       sequence-10212-e        7702 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 7702)
  TITLE     shMDC1 plasmids
REFERENCE   2  (bases 1 to 7702)
  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: pQCXIB CMV/TO DsRed2 (w444-1)
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1609..1912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1913..2116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2118..2136
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2139..2157
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2198..2222
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2260..2269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2266..2943
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2446..2466)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2858..2881
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(2966..2990)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3043..3084
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3131..3703
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3298..3315)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3525..3544
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3762..4160
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4408..4833
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4936..4958)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5094..5114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5099..5428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5279..5414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5450..5469
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5550..5567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5721..6309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5801..5820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6480..7271,7272..7340))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7103..7122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7341..7445)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7513..7531
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7631..7702,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg ataagcttgg gagttccgcg ttacataact
     1621 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1681 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1741 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1801 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1861 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1921 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1981 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2041 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2101 ctatataagc agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg
     2161 tcgactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagcagg
     2221 ctttaaagga accaattcag tcgactggat ccaccggtcg ccaccatggc ctcctccgag
     2281 aacgtcatca ccgagttcat gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac
     2341 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggccacaa caccgtgaag
     2401 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     2461 cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg
     2521 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggcg
     2581 accgtgaccc aggactcctc cctgcaggac ggctgcttca tctacaaggt gaagttcatc
     2641 ggcgtgaact tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc
     2701 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagaccca caaggccctg
     2761 aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag
     2821 cccgtgcagc tgcccggcta ctactacgtg gacgccaagc tggacatcac ctcccacaac
     2881 gaggactaca ccatcgtgga gcagtacgag cgcaccgagg gccgccacca cctgttcctg
     2941 tagcggccgc actcgagata tctagaccca gctttcttgt acaaagtggt tgatatccag
     3001 cacagtggcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     3061 cctctcctcg gtctcgattc tacgcgtacc ggtaggcctc gtacgcttaa ttaacggatc
     3121 cggaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     3181 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3241 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3301 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3361 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3421 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3481 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3541 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3601 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
     3661 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc acggggacgt
     3721 ggttttcctt tgaaaaacac gatgataagc ttgccacaac catggccaag cctttgtctc
     3781 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     3841 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     3901 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     3961 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     4021 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     4081 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     4141 gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc aggactgaca cgacccggac
     4201 gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag gcatctcatg agtgtgtctt
     4261 cccgttttcc gcctgaggtc actgcgtgga tggagcatca gtggtccagg ctctagtttt
     4321 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     4381 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     4441 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     4501 cagatcaagg tcaggaacag atggaacagg gtcgacccta gagaaccatc agatgtttcc
     4561 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4621 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4681 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     4741 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4801 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     4861 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg
     4921 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4981 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5041 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tagatccggc tgtggaatgt
     5101 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5161 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5221 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5281 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5341 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5401 cttttttgga ggcctaggct tttgcaaaaa gcttactggc ttaactatgc ggcatcagag
     5461 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5521 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6121 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6781 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7081 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7441 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7501 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt agcttggcca
     7561 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     7621 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     7681 gttcatagcc catatatgga gt
//
