LOCUS       sequence-10361-e        5589 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 5589)
  TITLE     shMDC1 plasmids
REFERENCE   2  (bases 1 to 5589)
  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: pLenti X1 Zeo/pTER shGFP #2 (733-1)
FEATURES             Location/Qualifiers
     source          1..5589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     814..831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    948..969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(984..989,990..1007,1008..1014)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1022..1038
                     /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     1027..1049
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1046..1062
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1046..1062
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1081..1101
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1083..1101
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             1357..1537
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1581..1706
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2199..2432
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2616..2660
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2921..3038
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    3070..3094
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3271..3290
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    3336..3354
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(3486..3510)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3541..4129
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3594..3614)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4012..4023)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4180..4200)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4185..4515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4366..4501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4527..4574
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4593..4967
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             5060..5293
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5365..5486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5402..5421)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5456..5475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      961 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1021 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     1081 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
     1141 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     1201 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     1261 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     1321 atattgtatt taagtgccta gctcgataca ataaacgggt ctctctggtt agaccagatc
     1381 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     1441 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1501 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacctga
     1561 aagcgaaagg gaaaccagag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     1621 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     1681 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     1741 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     1801 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1861 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1921 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1981 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     2041 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     2101 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     2161 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     2221 ttgggagcag caggaagcac tatgggcgca gcctcaatga cgctgacggt acaggccaga
     2281 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     2341 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     2401 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     2461 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     2521 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2581 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2641 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2701 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2761 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2821 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     2881 acagatccat tcgattagtg aacggatctc gacggttaac ttttaaaaga aaagggggga
     2941 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     3001 aagaattaca aaaacaaatt acaaaaattc aaaattttat tccagtgtgg tggaattctg
     3061 cagatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg
     3121 atccggtacc gaattcgaac gctgacgtca tcaacccgct ccaaggaatc gcggcccagt
     3181 gtcactaggc gggaacaccc agcgcgcgtg cgccctggca ggaagatggc tgtgagggac
     3241 aggggagtgg cgccctgcaa tatttgcatg tcgctatgtg ttctgggaaa tcaccataaa
     3301 cgtgaaatgt ctttggattt gggaatctta taagttccct atcagtgata gagatccccg
     3361 ctggagtaca actacaactt caagagagtt gtagttgtac tccagctttt tggaaaagct
     3421 tggtaccgag ctcggatcca ctagtccagt gtggtggaat tcgcggccgc actcgagata
     3481 tctagaccca gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcgac
     3541 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3601 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3661 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3721 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3781 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3841 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3901 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3961 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4021 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4081 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattctg
     4141 cagatatccg gttagtaatg agtttggaat taattctgtg gaatgtgtgt cagttagggt
     4201 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4321 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4441 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4501 ctaggctttt gcaaaaagct ccccctgttg acaattaatc atcggcatag tatatcggca
     4561 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     4621 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     4681 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     4741 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     4801 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     4861 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     4921 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4981 atttaaatgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5041 ttaaaagaaa aggggggact ggaagggcta gctcactccc aacgaagaca agatctgctt
     5101 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctgc
     5161 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5221 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5281 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     5341 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     5401 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5461 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     5521 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5581 ccgaggccg
//
