LOCUS       sequence-334367-        6522 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 6522)
  AUTHORS   Izon DJ, Punt JA, Xu L, Karnell FG, Allman D, Myung PS, Boerth NJ, 
            Pui JC, Koretzky GA, Pear WS
  TITLE     Notch1 regulates maturation of CD4+ and CD8+ thymocytes by 
            modulating TCR signal strength.
  JOURNAL   Immunity. 2001 Mar . 14(3):253-64.
  PUBMED    11290335
REFERENCE   2  (bases 1 to 6522)
  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: "Immunity. 
            2001 Mar . 14(3):253-64."
COMMENT     Sequence Label: NGFR
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(681..1472,1473..1541))
                     /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     1304..1323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1542..1646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1714..1732
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1770..1792)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1892..1911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2105..2127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2255..2274
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2474..2990
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3054..3395
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3462..3878
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3808..3830
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3846..3862
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3907..4474
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3924..4476
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4068..4085)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4295..4314
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             5390..5904
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6066..6082
                     /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(6090..6096,6097..6114,6115..6120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6135..6156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6273..6290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6444..6522,1..510))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
       61 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     1681 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1741 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1801 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1861 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1921 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1981 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2041 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     2101 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
     2161 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     2221 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     2281 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
     2341 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     2401 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     2461 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     2521 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     2581 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     2641 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     2701 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     2761 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     2821 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     2881 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
     2941 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
     3001 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
     3061 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     3121 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     3181 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     3241 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     3301 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     3361 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     3421 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     3481 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     3541 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     3601 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     3661 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     3721 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     3781 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     3841 cctcccttta tccagccctc actccttctc taggcgccgg aattagatct ctcgaggtta
     3901 acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     3961 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     4021 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     4081 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     4141 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     4201 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     4261 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     4321 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     4381 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     4441 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gacgggccgc
     4501 gcctgctgct gttgctgctt ctgggggtgt cccttggagg tgccaaggag gcatgcccca
     4561 caggcctgta cacacacagc ggtgagtgct gcaaagcctg caacctgggc gagggtgtgg
     4621 cccagccttg tggagccaac cagaccgtgt gtgagccctg cctggacagc gtgacgttct
     4681 ccgacgtggt gagcgcgacc gagccgtgca agccgtgcac cgagtgcgtg gggctccaga
     4741 gcatgtcggc gccgtgcgtg gaggccgacg acgccgtgtg ccgctgcgcc tacggctact
     4801 accaggatga gacgactggg cgctgcgagg cgtgccgcgt gtgcgaggcg ggctcgggcc
     4861 tcgtgttctc ctgccaggac aagcagaaca ccgtgtgcga ggagtgcccc gacggcacgt
     4921 attccgacga ggccaaccac gtggacccgt gcctgccctg caccgtgtgc gaggacaccg
     4981 agcgccagct ccgcgagtgc acacgctggg ccgacgccga gtgcgaggag atccctggcc
     5041 gttggattac acggtccaca cccccagagg gctcggacag cacagccccc agcacccagg
     5101 agcctgaggc acctccagaa caagacctca tagccagcac ggtggcaggt gtggtgacca
     5161 cagtgatggg cagctcccag cccgtggtga cccgaggcac caccgacaac ctcatccctg
     5221 tctattgctc catcctggct gctgtggttg tgggccttgt ggcctacata gccttcaaga
     5281 ggtggaacag tcatcgatat cctcgaggtc accgcggtct agagtcgacc tgcagccaag
     5341 cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     5401 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     5461 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     5521 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     5581 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     5641 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5701 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     5761 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     5821 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5881 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     5941 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     6001 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     6061 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac ta
//
