LOCUS       sequence-124047-        6265 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 6265)
  AUTHORS   Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, Liu A, 
            Miu K, Watnick RS, Reinhardt F, McAllister SS, Jacks T, Weinberg RA
  TITLE     Growth-inhibitory and tumor- suppressive functions of p53 depend on 
            its repression of CD44 expression.
  JOURNAL   Cell. 2008 Jul 11. 134(1):62-73.
  PUBMED    18614011
REFERENCE   2  (bases 1 to 6265)
  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: "Cell. 2008 
            Jul 11. 134(1):62-73."
COMMENT     Sequence Label: shCD44-3 pRRL
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(130..198,199..990)
                     /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(348..367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1161..1749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1650..1669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2370..2603
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2788..2832
                     /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"
     promoter        3130..3378
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3130..3150
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3301..3320
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    3476..3593
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3642..4152
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4071..4089
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      4168..4177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4174..4176,4177..4179,4180..4893)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4174..4176,4177..4179,4180..4890)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4219..4240)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4480..4499)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4827..4848
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5001..5234
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5306..5427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5343..5362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5397..5416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5467..5602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5622..5641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5623..5641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5651..5668)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5651..5667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5660..5682)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5809..6237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5896..5915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6106..6127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
       61 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      121 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      181 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      241 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      301 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      361 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      421 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
      481 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
      541 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
      601 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
      661 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
      721 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
      781 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
      841 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
      901 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
      961 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1021 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1081 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1141 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1201 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1261 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1321 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1381 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1441 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1501 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1561 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1621 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1681 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1741 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1801 gctcacatgt tctttcctgc gttatcccct gattctctag cggaggctag aaggagagag
     1861 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1921 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1981 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     2041 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     2101 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     2161 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     2221 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2281 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2341 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2401 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2461 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2521 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2581 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2641 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2701 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2761 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2821 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2881 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2941 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     3001 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     3061 attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc
     3121 cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     3181 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
     3241 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     3301 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     3361 tggaaaggac gaaacaccgg ttgtaacacc tacaccatta tcctcgagga taatggtgta
     3421 ggtgttacat ttttgaattc tcgacctcga gacaaatggc agtattcatc cacaatttta
     3481 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     3541 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     3601 attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt
     3661 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     3721 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     3781 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     3841 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     3901 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     3961 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     4021 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     4081 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     4141 ctctctcccc agggggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     4201 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4261 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4321 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4381 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4441 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     4501 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     4561 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     4621 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     4681 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     4741 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     4801 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4861 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgtcgacgg gcccgcggaa
     4921 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     4981 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     5041 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5101 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5161 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5221 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     5281 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     5341 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5401 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     5461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5521 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5581 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     5641 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5701 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5761 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     5821 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5881 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5941 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6001 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6061 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6121 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6181 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6241 acaaaatatt aacgcttaca attta
//
