LOCUS       sequence-9636-ex        3783 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 3783)
  AUTHORS   Beard C, Hochedlinger K, Plath K, Wutz A, Jaenisch R
  TITLE     Efficient method to generate single-copy transgenic mice by 
            site-specific integration in embryonic stem cells.
  JOURNAL   Genesis. 2006 Jan . 44(1):23-8.
  PUBMED    16400644
REFERENCE   2  (bases 1 to 3783)
  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: "Genesis. 
            2006 Jan . 44(1):23-8."
COMMENT     Sequence Label: Col1a-3'probe
FEATURES             Location/Qualifiers
     source          1..3783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(892..908)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(893..909)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(938..958)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(938..956)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(977..993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(977..993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(990..1012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1001..1017
                     /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(1025..1031,1032..1049,1050..1055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1070..1091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1208..1225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1379..1967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1459..1478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2138..2929,2930..2998))
                     /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     2761..2780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2999..3103)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3129..3584)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3266..3287)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3478..3497
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3711..3733
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3725..3742
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3726..3742
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3752..3771
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3752..3770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccaccgcggt ggcggccgct ctagataaat tttcctttca cgttgtcctc atgcaaagtg
       61 ctaataataa attattaaag gccatcatta atagtcttgg atactccgtg accatatgaa
      121 cgtgtcctat gggtgccaca gtagactggt agcacaagag agggaagcta caaatggcct
      181 gttccttcct ttcaaatttg agacctgtct ccccgagtga aaggggtcat ctgtgtcctt
      241 atgtctactg tctggttcac cccagacttc tgcctgctgg attctcgggc cttctactat
      301 gtcaccgtcc ccattctctg agatgtgggg gacaattagt taccatccct ctagggcctc
      361 aggttctcag atggacagag ggagagaggg aaaaggaaga agaaaaagca aaaagttttg
      421 gggataaagg aaaaggaaag aagggggcag gaagaagggg ggaacaattc cacgggctcc
      481 ctgtaccgag ccagcatgct atcgtcccgg gcacctctgt gatgtggggt cctgtccttt
      541 ctggtgacac cccggaagtt ccaccacagc tctctaagat cccccgggga gagttccttg
      601 agggctgggc acctctctct ggtgagatca agggtgaccc aagaaattct catcctcctt
      661 gtgacacccc agcattagaa ggccccttct atactaaatt atggtttctt tgggctagag
      721 gtggggagag gccagctacc cctccatgtg tgaccaagga gcaacctggt cctccatgtc
      781 tcagtccctg tttctgctgc ttgaatccct ttgagggctc atgaacctcc caggatccat
      841 gggcaacacc tcccaggcct caggtctgca ggaattcgat atcaagctta tcgataccgt
      901 cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
      961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1801 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3121 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3181 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3241 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3301 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3361 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3421 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3481 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3541 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     3601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     3661 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     3721 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattgga
     3781 gct
//
