LOCUS       sequence-162798-        5818 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 5818)
  AUTHORS   Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C, 
            Savagner P, Gitelman I, Richardson A, Weinberg RA
  TITLE     Twist, a master regulator of morphogenesis, plays an essential role 
            in tumor metastasis.
  JOURNAL   Cell 2004 Jun 25;117(7):927-39.
  PUBMED    15210113
REFERENCE   2  (bases 1 to 5818)
  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"; 
            date: "2004-06-25"; volume: "117(7)"; pages: "927-39"
COMMENT     Sequence Label: pBABE-puro-mTwist
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             68..537
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(758..1357)
                     /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(841..861)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1338..1357
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1367..1696)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1381..1516
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1681..1701
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(2491..2907)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2507..2523)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2539..2561)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3108..3307
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3372..3841
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3872..3976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3977..4045,4046..4837)
                     /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"
     rep_origin      4997..5585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5486..5505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5739..5756
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caactatggc catttaatgt aaatacttaa gaaaaaaaac caaattaatt ttgatacatg
       61 ctgcatgtga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      121 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      181 gggcgactca gtcaatcgga ggactggcgc cccgagtgag gggttgtggg ctcttttatt
      241 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      301 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      361 gaaactgctg agggctggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      421 aggaactgct taccacagat atcctgtttg gcccatattc agctgttcca tctgttcttg
      481 gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca tattcagctg
      541 ttccatctgt tcctgacctt gatctgaact tctctattct cagttatgta tttttccatg
      601 ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc
      661 cccccttttt ctggagacta aataaaatct tttattttat cgatgcatgg ggtcgtgcgc
      721 tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca
      781 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
      841 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
      901 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
      961 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     1021 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     1081 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     1141 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     1201 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     1261 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     1321 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt aagctttttg caaaagccta
     1381 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     1441 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     1501 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     1561 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     1621 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     1681 accctaactg acacacattc cacagggtcg accactgtgc tggcgaattc ctacccgctg
     1741 ctagtgggac gcggacatgg accaggcccc ctccatcctc cagacggaga aggcgtagct
     1801 gagccgctcg tgggccacat agctgcagct tgccatcttg gagtccagct cgtcgctctg
     1861 caggacctgg tacaggaagt cgatgtacct ggccgccagt ttgagggtct gaatcttgct
     1921 cagcttgtcc gagggcagcg tggggatgat cttgcgcagg gcggcgaacg cctcgttcag
     1981 cgactgcgtg cgctggcgct cccgcacgtt agccatgacc cgctgggtct gcagctcctc
     2041 gtacgactgc gggctcccgc ccccgctgct gctgccgccg ccgccgccac cacctccgcc
     2101 cgcgccgccg ccgccgcctc cgcccgcaga tttcttgccg cgcttgccct gggccgggct
     2161 gcctggctcg tcgccgcctc cgatgccccc gcccgtggcc ccgccgggcc ccgcgctgcc
     2221 gcccgcgctg cgccgactgc tgcgtctctt gcgagccccg cgcttgccgc tcgccggctg
     2281 ctgccggtcc ggctcctcct cgctgttgct caggctgtcg tcggccggag agactggcga
     2341 gctggacacg tcctgcatca tctcgcgggc ggcgacgtgc ggcctcgcgg agccgggacg
     2401 ggggagggta gagcagagga gcggggcgcc ctagcccgcc agcttccccg cgctcggcgc
     2461 cggcccgtac caccacactg ggatccggcc ggcgcctaga gaaggagtga gggctggata
     2521 aagggaggat cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg
     2581 aggaagagga ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa
     2641 agccaaggct tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg
     2701 tgaaaagacc ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg
     2761 ccatccgacg ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt
     2821 gacatctacc gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa
     2881 acttaaggga gtggtaacag tctggcccta attttcagac aaatacagaa acacagtcag
     2941 acagagacaa cacagaacga tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt
     3001 ccaaaccgaa agcaaaaatt cagacggagg cgggaactgt tttaggttct cgtctcctac
     3061 cagaaccaca tatcctgacg gggtcggatt ccacatcgac tcccttcctc aggtcgggcc
     3121 acaaaaacgg cccccaaagt ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa
     3181 ctcgtcagtt ccaccacggg tccgccagat acagagctag ttagctaact agtaccgacg
     3241 caggcgcata aaatcagtca tagacactag acaatcggac agacacagat aagttgctgg
     3301 ccagcttgcc tcccggtggt gggtcggtgg tccctgggca ggggtctccc gatcccggac
     3361 gagcccccaa atgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc
     3421 aaggaacagc gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt
     3481 acccgggcga ctcagtcaat cggaggactg gcgccccgag tgaggggttg tgggctcttt
     3541 tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt
     3601 tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc
     3661 tctagaaact gctgagggct ggaccgcatc tggggaccat ctgttcttgg ccctgagccg
     3721 gggcaggaac tgcttaccac agatatcctg tttggcccat attcagctgt tccatctgtt
     3781 cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg cccatattca
     3841 ggctgcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3961 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4021 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4081 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4141 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4201 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4261 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4321 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4381 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4441 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4501 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4561 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4621 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4681 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4741 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4801 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4861 tatactttag attgatttgc ggccgcaaaa ggatctaggt gaagatcctt tttgataatc
     4921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatg
//
