LOCUS       sequence-9637-ex        3335 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 3335)
  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 3335)
  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-5'probe
FEATURES             Location/Qualifiers
     source          1..3335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..36
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     20..36
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(490..510)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(490..508)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(529..545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(529..545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(542..564)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    553..569
                     /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(577..583,584..601,602..607))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    622..643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(760..777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(931..1519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1011..1030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1690..2481,2482..2550))
                     /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     2313..2332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2551..2655)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2681..3136)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2818..2839)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3030..3049
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3263..3285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3277..3294
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3278..3294
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3304..3323
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3304..3322
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg cccccgggta
       61 agttgcatct tccattactt cttcctgggt tcctccctgt cttggttact ttcagatatc
      121 cctctgttct ccagagaaga gttcagagac cagacagggg gtggggagaa aaagaagctt
      181 ggtcttacag aacctttgac cttcccaccc gtgccccctt gcaaacctgg ccataggaac
      241 tagttgaggg caccctctga gagacagaag cagagactgg gcggagccag cagtgggggg
      301 aggggaaaca cttctgagcc ggagtttggg ggctgccttg agtttctagc cactttttct
      361 aaccatctct ctatccctag ggtccccctg gctctgctgg ttctcctggc aaagacggac
      421 tcaacggtct ccctggcccc attggtcccc ctggtcctcg agggggggcc cggtacccag
      481 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
      541 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      601 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      661 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      721 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      781 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      841 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     1381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1741 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1801 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1861 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1921 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1981 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2041 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2101 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2161 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2221 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2281 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2341 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2401 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2461 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2521 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2581 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2641 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     2701 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2761 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2821 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2881 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2941 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3001 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3061 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3121 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     3301 gcgtaatacg actcactata gggcgaattg gagct
//
