LOCUS       sequence-127693-        5875 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 5875)
  AUTHORS   Viswanathan SR, Powers JT, Einhorn W, Hoshida Y, Ng TL, Toffanin S, 
            O'Sullivan M, Lu J, Phillips LA, Lockhart VL, Shah SP, Tanwar PS, 
            Mermel CH, Beroukhim R, Azam M, Teixeira J, Meyerson M, Hughes TP, 
            Llovet JM, Radich J, Mullighan CG, Golub TR, Sorensen PH, Daley GQ
  TITLE     Lin28 promotes transformation and is associated with advanced human 
            malignancies.
  JOURNAL   Nat Genet. 2009 Jul . 41(7):843-8.
  PUBMED    19483683
REFERENCE   2  (bases 1 to 5875)
  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: "Nat Genet. 
            2009 Jul . 41(7):843-8."
COMMENT     Sequence Label: pBABE-hLin28B
FEATURES             Location/Qualifiers
     source          1..5875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(810..1409)
                     /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(893..913)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1390..1409
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1419..1748)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1433..1568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1733..1753
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(2560..2583)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2600..3016)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2616..2632)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2648..2670)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3217..3416
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3481..3950
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3981..4085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4086..4154,4155..4946)
                     /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(4304..4323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5106..5694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5595..5614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5848..5865
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
      121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
      241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
      301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
      541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
      601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
      661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
      721 ttctggagac taaataaaat cttttatttt atcgatgcat ggggtcgtgc gctcctttcg
      781 gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag
      841 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
      901 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
      961 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     1021 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     1081 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     1141 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     1201 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     1261 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     1321 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     1381 aggcgcaccg tgggcttgta ctcggtcatg gtaagctttt tgcaaaagcc taggcctcca
     1441 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     1501 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1561 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1621 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     1681 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     1741 tgacacacat tccacagggt cgacttatgt ctttttcctt ttttgaactg aaggcccctt
     1801 tttgctttgc tcttctggtg ctatagatga cttcgtggag gaagcttctt gaggtgacct
     1861 gcctgaccgt tctgagatct ctgccctagc ctcctgagga aacggtggtg atgtacagcc
     1921 atgcccgcct cccacttctc gagggagagt tgaagtgcat ggctgggatt ctgcttcctg
     1981 tcttccctga gaactcgcgg gtggctgtgc aacattttta tgtgggcagt ttgccaccat
     2041 gtgcatgatg ctctgacagt aatggcactt ctttggctga ggaggtagac tacattcctt
     2101 agcatgatga tcaaggccac cacagttgta gcatctatct ccctttggtt ttcttttctg
     2161 tagtgtcttc cctttgggtc ttctttcact tcctaaacag gggctcccac caggtcctgt
     2221 tacccgtatt gactcaaggc ctttggaaga ttttttaaat gtgaattcca ctggttctcc
     2281 ttcttttagg cttctaaatc cttccatgaa tagtttgctt tggtgtacaa atacatcgac
     2341 tggaatatcc aaggggcttc cctctcggtt tatcatggag atgaatccaa atcccatgcg
     2401 cacattgaac cacttacagt ggccagttcc gcgcaaaacc tgggattcct cctctgccgg
     2461 ctccggcagc ttcccgggct cttctccacc acctttgcta gccccgcctt cggccattgg
     2521 taccgatatc agatctatcg atgaattcgc ggccgcaagc ttgtcgtcat cgtctttgta
     2581 gtccatggtg gatccggccg gcgcctagag aaggagtgag ggctggataa agggaggatc
     2641 gaggcggggt cgaacgagga ggttcaaggg ggagagacgg ggcggatgga ggaagaggag
     2701 gcggaggctt agggtgtaca aagggcttga cccagggagg ggggtcaaaa gccaaggctt
     2761 cccaggtcac gatgtagggg acctggtctg ggtgtccatg cgggccaggt gaaaagacct
     2821 tgatcttaac ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc catccgacgt
     2881 taaaggttgg ccattctgca gagcagaagg taacccaacg tctcttcttg acatctaccg
     2941 actggttgtg agcgatccgc tcgacatctt tccagtgacc taaggtcaaa cttaagggag
     3001 tggtaacagt ctggccctaa ttttcagaca aatacagaaa cacagtcaga cagagacaac
     3061 acagaacgat gctgcagcgc tgcagcagac aagacgcgcg gcttcggttc caaaccgaaa
     3121 gcaaaaattc agacggaggc gggaactgtt ttaggttctc gtctcctacc agaaccacat
     3181 atcctgacgg ggtcggattc cacatcgact cccttcctca ggtcgggcca caaaaacggc
     3241 ccccaaagtc cctgggacgt ctcccagggt tgcggccggg tgttcagaac tcgtcagttc
     3301 caccacgggt ccgccagata cagagctagt tagctaacta gtaccgacgc aggcgcataa
     3361 aatcagtcat agacactaga caatcggaca gacacagata agttgctggc cagcttgcct
     3421 cccggtggtg ggtcggtggt ccctgggcag gggtctcccg atcccggacg agcccccaaa
     3481 tgaaagaccc ccgctgacgg gtagtcaatc actcagagga gaccctccca aggaacagcg
     3541 agaccacaag tcggatgcaa ctgcaagagg gtttattgga tacacgggta cccgggcgac
     3601 tcagtcaatc ggaggactgg cgccccgagt gaggggttgt gggctctttt attgagctcg
     3661 gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc
     3721 acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg
     3781 ctgagggctg gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact
     3841 gcttaccaca gatatcctgt ttggcccata ttcagctgtt ccatctgttc ttggccctga
     3901 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag gctgcaggtg
     3961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4021 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4081 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4141 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4201 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4261 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4321 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4381 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4441 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4501 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4561 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4621 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4681 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4741 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4801 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4861 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4921 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4981 ttgatttgcg gccgcaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5221 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5761 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5821 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
