LOCUS       sequence-62293-e        4184 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 4184)
  AUTHORS   Singer PA, Trevor K, Oshima RG
  TITLE     Molecular cloning and characterization of the Endo B cytokeratin 
            expressed in preimplantation mouse embryos.
  JOURNAL   J Biol Chem. 1986 Jan 15;261(2):538-47.
  PUBMED    2416755
REFERENCE   2  (bases 1 to 4184)
  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: "J Biol 
            Chem."; date: "1986-01-15"; volume: "261(2)"; pages: "538-47"
COMMENT     Sequence Label: PUC9B71A
FEATURES             Location/Qualifiers
     source          1..4184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1551..1568)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1551..1567)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1560..1582)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1776..1795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1895..1917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1955..1973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2041..2145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2146..2214,2215..3006)
                     /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(2364..2383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3177..3765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3666..3685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3919..3936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4053..4074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4089..4094,4095..4112,4113..4119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4127..4143
                     /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)."
     primer_bind     4132..4154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4151..4167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4151..4167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agcttggctg cacccccccc ccccccctcc gcggcggaac tcctgttctg gtctctcgct
       61 tcgctctcct ctccagacaa gatgagcttc acaactcgct ccaccacctt ctccaccaac
      121 taccggtccc tgggctctgt gcgaactccc agccagcggg tccggcctgc cagcagcgca
      181 gccagcgtct atgcaggtgc tgggggctcc gggtcccgga tatccgtgtc ccgctctgtc
      241 tggggtggct ctgtggggtc cgcaggcctg gcgggaatgg gtggaatcca gaccgagaaa
      301 gagaccatgc aagacctgaa cgatcgcctg gccagctacc tagacaaggt gaagagcctg
      361 gaaactgaga acaggagact ggagagcaaa atccgggaac atctggagaa gaaggggccc
      421 cagggcgtca gagactgggg ccactacttc aagatcatcg aagacctgag ggctcagatc
      481 cttgcgaatt ctgtggacaa tgcccgcatc gtcttgcaga tcgacaatgc ccgccttgcc
      541 gccgatgact ttagagtcaa gtatgagaca gaactagcca tgcgccagtc tgtggagagc
      601 gacatccatg gactccgcaa ggtggtagat gacaccaaca tcacaaggct gcagctggag
      661 acagaaatcg aggcactcaa ggaagaactt ctgttcatga agaagaatca tgaagaggaa
      721 gtccaaggtc tggaagccca gattgccagc tctggattga ctgtggaagt ggatgccccc
      781 aaatctcagg acctcagcaa gatcatggcg gacatccgcg cccagtatga agcgctggct
      841 cagaagaacc gcgaggaact ggacaagtac tggtctcagc agattgagga gagtaccaca
      901 gttgtcacca ccaagtctgc cgaaatcagg gacgctgaga ccacactcac ggagctgaga
      961 cgcaccctcc agaccttgga gattgacttg gactccatga aaaaccagaa catcaacttg
     1021 gagaacagcc tcggggatgt ggaggcccga tacaaggcac agatggagca gctcaatggg
     1081 gtccttctgc atctggagtc agagctggca caaactcggg cagagggcca gcgccaggcc
     1141 caggaatatg aagccctctt gaacatcaag gtgaagcttg aggcagagat tgccacctac
     1201 cgccgcttgc tggaggatgg agaagatttc agtctcaacg atgccctgga ctccagcaac
     1261 tccatgcaaa ctgtgcagaa gacaactacc cgtaagatcg tggatggcag agtggtgtcc
     1321 gagactaatg acaccagagt tctgaggcac tgaggcagag aaggagggaa cccctgggaa
     1381 ctgagggacc aataaaagtt gagagctcac tggaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1441 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1501 aaaaaaaagg gggggggggg gggtgcaggt cgacggatcc ccgggaattc actggccgtc
     1561 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     1621 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1681 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     1741 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     1801 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     1861 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     1921 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     1981 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3961 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4021 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4081 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4141 caatttcaca caggaaacag ctatgaccat gattacgcca agct
//
