LOCUS       sequence-129555-        6376 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 6376)
  AUTHORS   Padua D, Zhang XH, Wang Q, Nadal C, Gerald WL, Gomis RR, Massague J
  TITLE     TGFbeta primes breast tumors for lung metastasis seeding through 
            angiopoietin-like 4.
  JOURNAL   Cell. 2008 Apr 4. 133(1):66-77.
  PUBMED    18394990
REFERENCE   2  (bases 1 to 6376)
  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 
            Apr 4. 133(1):66-77."
COMMENT     Sequence Label: pRetroSuper shAngptl4 puro
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     CDS             complement(783..1382)
                     /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(866..886)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1363..1382
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1407..1906)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1455..1474)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1848..1871
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1917..2131
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2052..2071
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(2204..2220)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2221)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(2239..2655)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2255..2271)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2287..2309)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2722..3063
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3127..3643
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3843..3862)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3990..4012)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4206..4225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4325..4347
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4385..4403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4471..4575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4576..4644,4645..5436)
                     /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(4794..4813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5607..6195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6096..6115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6349..6366
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
      241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
      301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
      361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
      481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
      541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagcttgcca
      661 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
      721 tttatcgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc gtggggcggg
      781 cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
      841 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
      901 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
      961 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     1021 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     1081 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     1141 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     1201 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     1261 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     1321 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     1381 atggtaagct agcttgggct gcaggtcgaa aggcccggag atgaggaaga ggagaacagc
     1441 gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga ccttcgggcg
     1501 cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct tcccagcctc
     1561 tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag gcctacccgc
     1621 ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc tacttccatt
     1681 tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac taggggagga
     1741 gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct accggtggat
     1801 gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag cggggctgct
     1861 aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggta gaattcgaac
     1921 gctgacgtca tcaacccgct ccaaggaatc gcgggcccag tgtcactagg cgggaacacc
     1981 cagcgcgcgt gcgccctggc aggaagatgg ctgtgaggga caggggagtg gcgccctgca
     2041 atatttgcat gtcgctatgt gttctgggaa atcaccataa acgtgaaatg tctttggatt
     2101 tgggaatctt ataagttctg tatgagacca cagatccccc agcaggatcc agcaactctt
     2161 caagagagag ttgctggatc ctgctgtttt tggaaaagct tatcgatacc gtcgacctcg
     2221 agagatcgat ctaattccgg cgcctagaga aggagtgagg gctggataaa gggaggatcg
     2281 aggcggggtc gaacgaggag gttcaagggg gagagacggg gcggatggag gaagaggagg
     2341 cggaggctta gggtgtacaa agggcttgac ccagggaggg gggtcaaaag ccaaggcttc
     2401 ccaggtcacg atgtagggga cctggtctgg gtgtccatgc gggccaggtg aaaagacctt
     2461 gatcttaacc tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt
     2521 aaaggttggc cattctgcag agcagaaggt aacccaacgt ctcttcttga catctaccga
     2581 ctggttgtga gcgatccgct cgacatcttt ccagtgacct aaggtcaaac ttaagggagt
     2641 ggtaacagtc tggccctaat tttcagacaa atacagaaac acagtcagac agagacaaca
     2701 cagaacgatg ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag
     2761 caaaaattca gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata
     2821 tcctgacggg gtcggattcc acatcgactc ccttcctcag gtcgggccac aaaaacggcc
     2881 cccaaagtcc ctgggacgtc tcccagggtt gcggccgggt gttcagaact cgtcagttcc
     2941 accacgggtc cgccagatac agagctagtt agctaactag tacagacgca ggcgcaaaca
     3001 ttagatgccg gcacaaacac gcacaaagac agagacagac acgaaacgac cgctggccag
     3061 cttacctccc ggcggggggg tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa
     3121 cctccaaatg aaagaccccc gaggtgggca gtcaatcaat ctgaggagac cctcccaagg
     3181 atcagcgagt ccacgattcg gatgcaaaca gcaagaggct ttattgggaa tacgggtacc
     3241 cgggcgacgc agtctatcgg aggactggcg cgccgagtga ggggttgtgg gctcttttat
     3301 tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca
     3361 aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct
     3421 agaaactgct gagggcggga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg
     3481 caggaactgc ttaccacaga tatcctgttt ggcccatatt ctgctgtctc tctgttccta
     3541 accttgatct gaacttctct attctcagtt atgtattttc catgccttgc aaaatggcgt
     3601 tacttaagct agcttgccaa acctacaggt ggggtctttc attcccccct ttttctggag
     3661 actaaataaa atcttttatt ttatctatgg ctcgtactct ataggcttca gctggtgata
     3721 ttgttgagtc aaaactagag cctggaccac tgatatcctg tctttaacaa attggactaa
     3781 tcgcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     3841 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     3901 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     3961 ctccttgcat gcaccattcc ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct
     4021 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4081 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     4141 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     4201 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4261 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4321 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4381 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4441 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4741 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6241 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6301 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6361 agcgaggaag cggaag
//
