LOCUS       sequence-140265-        6468 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 6468)
  AUTHORS   He W, Dorn DC, Erdjument-Bromage H, Tempst P, Moore MA, Massague J
  TITLE     Hematopoiesis controlled by distinct TIF1gamma and Smad4 branches of
            the TGFbeta pathway.
  JOURNAL   Cell. 2006 Jun 2. 125(5):929-41.
  PUBMED    16751102
REFERENCE   2  (bases 1 to 6468)
  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. 2006 
            Jun 2. 125(5):929-41."
COMMENT     Sequence Label: pRetroSuper-GFP Smad2
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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(join(695..1408,1409..1411,1412..1414))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(740..761)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1089..1108
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1348..1369
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1411..1420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1451..1950)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1499..1518)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1892..1915
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1961..2175
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2096..2115
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(2248..2264)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2249..2265)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(2279..2695)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2295..2311)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2327..2349)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2762..3103
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3167..3683
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3883..3902)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4030..4052)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4246..4265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4365..4387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4425..4443)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4511..4615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4616..4684,4685..5476)
                     /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(4834..4853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5647..6235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6389..6406
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa
      661 tcttttattt tatcgataat tcttaattaa ttcattactt gtacagctcg tccatgccga
      721 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
      781 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
      841 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
      901 tgttgtggcg gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg gccatgatat
      961 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     1021 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     1081 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     1141 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     1201 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     1261 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     1321 cagacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     1381 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gttaattaag aattagcttg
     1441 ggctgcaggt cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct
     1501 tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg ggcgcccgcc ccgcccctga
     1561 gcccgcccct gagcccgccc ccggacccac cccttcccag cctctgagcc cagaaagcga
     1621 aggagcaaag ctgctattgg ccgctgcccc aaaggcctac ccgcttccat tgctcagcgg
     1681 tgctgtccat ctgcacgaga ctagtgagac gtgctacttc catttgtcac gtcctgcacg
     1741 acgcgagctg cggggcgggg gggaacttcc tgactagggg aggagtagaa ggtggcgcga
     1801 aggggccacc aaagaacgga gccggttggc gcctaccggt ggatgtggaa tgtgtgcgag
     1861 gccagaggcc acttgtgtag cgccaagtgc ccagcggggc tgctaaagcg catgctccag
     1921 actgccttgg gaaaagcgcc tcccctaccc ggtagaattc gaacgctgac gtcatcaacc
     1981 cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa cacccagcgc gcgtgcgccc
     2041 tggcaggaag atggctgtga gggacagggg agtggcgccc tgcaatattt gcatgtcgct
     2101 atgtgttctg ggaaatcacc ataaacgtga aatgtctttg gatttgggaa tcttataagt
     2161 tctgtatgag accacagatc cccggattgc cacatgttat atttcaagag aatataacat
     2221 gtggcaatcc tttttggaaa agcttatcga taccgtcgac ctcgagagat ctaattccgg
     2281 cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag
     2341 gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa
     2401 agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga
     2461 cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga
     2521 ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag
     2581 agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct
     2641 cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggccctaat
     2701 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcgct
     2761 gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca gacggaggcg
     2821 ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg gtcggattcc
     2881 acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc ctgggacgtc
     2941 tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc cgccagatac
     3001 agagctagtt agctaactag tacagacgca ggcgcaaaca ttagatgccg gcacaaacac
     3061 gcacaaagac agagacagac acgaaacgac cgctggccag cttacctccc ggcggggggg
     3121 tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa cctccaaatg aaagaccccc
     3181 gaggtgggca gtcaatcaat ctgaggagac cctcccaagg atcagcgagt ccacgattcg
     3241 gatgcaaaca gcaagaggct ttattgggaa tacgggtacc cgggcgacgc agtctatcgg
     3301 aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
     3361 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
     3421 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggcggga
     3481 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     3541 tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct gaacttctct
     3601 attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct agcttgccaa
     3661 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
     3721 ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc aaaactagag
     3781 cctggaccac tgatatcctg tctttaacaa attggactaa tcgcctctac gccggacgca
     3841 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     3901 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     3961 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     4021 ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4081 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4141 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     4201 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     4261 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4321 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4381 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4441 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4501 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4621 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4681 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4741 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4801 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4861 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4921 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4981 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5041 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5101 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5161 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5221 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5281 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5341 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5401 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5461 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6301 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6361 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6421 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatg
//
