LOCUS       sequence-434418-        5934 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 5934)
  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 5934)
  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: pCMV-FLAG TIF1gamma M (456-887)
FEATURES             Location/Qualifiers
     source          1..5934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             619..642
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1112..1123)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1979..2601
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2178..2197)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2625..2645)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2790..2924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2976..2995)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2977..2995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3009..3025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3009..3025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3022..3044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3033..3049
                     /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)."
     promoter        complement(join(3057..3063,3064..3081,3082..3087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3102..3123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3240..3257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3411..3999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4170..4961,4962..5030))
                     /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     4793..4812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5031..5135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5162..5617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5249..5268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5459..5480
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5743..5765
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5757..5774
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5758..5774
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgcag tgaccatgga ctacaaggac gacgatgaca aggaattcag atccaccatg
      661 gcaatacgtt tccattgtga tcccaccttc tgggcaaaga atgtagtcaa tttaggtaat
      721 ctagtaatag agagtaaacc agctcctggt tatactccta atgttgtagt tgggcaagtt
      781 cctccaggga caaaccacat tagtaaaacc cctggacaga ttaacttagc acagcttcga
      841 ctccagcaca tgcaacaaca agtatatgca cagaaacatc agcagttgca acagatgagg
      901 atgcagcaac caccagcacc tgtaccaact acaacaacaa caacacaaca gcatcctaga
      961 caagcagccc ctcagatgtt acaacaacag cctcctcgat tgatcagtgt gcaaacaatg
     1021 caaagaggca acatgaactg tggagctttt caagcccatc agatgagact ggctcagaat
     1081 gctgccagaa taccagggat acccaggcac agcggccctc aatattccat gatgcagcca
     1141 cacctccaaa gacaacactc aaacccaggg catgctggac cctttcccgt agtatcggta
     1201 cacaacacca caatcaaccc aacgagccct actacagcaa ctatggcaaa tgcaaaccga
     1261 ggtcccacca gcccatctgt tacagcaata gagctaatcc cctcagttac caatccagaa
     1321 aaccttccat cgctgccaga tattccaccc atacagttgg aagatgctgg ctcaagtagt
     1381 ttagataatc tactaagtag atacatctca ggcagtcacc tacccccaca gcctacaagc
     1441 accatgaatc cttctccagg tccctctgcc ctttctccgg gatcatcagg tttatccaat
     1501 tctcacacac ctgtgagacc cccaagtact tctagtactg gcagtcgagg cagctgtggg
     1561 tcatcaggaa gaactgctga gaagacaagt cttagtttca aatctgatca ggtgaaggtc
     1621 aagcaagaac ctgggactga agatgaaata tgtagctttt caggaggtgt aaaacaagaa
     1681 aaaacagagg atggcaggag gagtgcttgc atgttgagca gtcctgagag tagcttgaca
     1741 ccacctctct caaccaacct gcatctagaa agtgaattgg atgcattggc aagcctggaa
     1801 aaccatgtga aaactgaacc tgcagatatg aatgaaagct gcaaacagtc agggctcagc
     1861 agccttgtta atggaaagtc cccaattcga agcctcatgc acaggtcggc aaggattgga
     1921 ggagatggca acaataaaga tgatgaccca aatgaatgac tcgactctag aggatcccgg
     1981 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2041 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2101 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2161 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2221 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2281 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2341 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     2401 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     2461 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     2521 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     2581 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     2641 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2701 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2761 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca gttccgccca
     2821 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2881 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2941 gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc
     3001 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3061 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3121 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3181 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5161 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5221 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5281 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5341 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5401 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5461 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5521 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5581 aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca
     5641 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5701 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     5761 aaacgacggc cagtgccaag ctgatctata cattgaatca atattggcaa ttagccatat
     5821 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     5881 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgtt
//
