LOCUS       sequence-14446-e        4133 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 4133)
  AUTHORS   Dmitriev PV, Vassetzky YS
  TITLE     A set of vectors for introduction of antibiotic resistance genes by 
            in vitro Cre-mediated recombination.
  JOURNAL   BMC Res Notes. 2008 Dec 23;1:135. doi: 10.1186/1756-0500-1-135.
  PUBMED    19105832
REFERENCE   2  (bases 1 to 4133)
  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: "BMC Res 
            Notes."; date: "2008-12-23"; volume: "1:135. doi"; pages: " 
            10.1186/1756-0500-1-135"
COMMENT     Sequence Label: pT-TK
FEATURES             Location/Qualifiers
     source          1..4133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..759
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          829..961
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     944..963
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1006..1024
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1028..1037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1034..1969
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2000..2121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2011..2030)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2065..2084
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2254..2358
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2373..2406
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2448..3239,3240..3308))
                     /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     3071..3090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3478..4066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3967..3986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
       61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
      121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
      181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
      241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
      301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
      361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
      421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
      481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
      541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
      601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
      661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
      721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca
      781 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      841 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      901 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      961 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
     1021 taggctagcc accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac
     1081 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta
     1141 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc
     1201 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc
     1261 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga
     1321 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt
     1381 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa
     1441 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg
     1501 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct
     1561 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc
     1621 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac
     1681 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat
     1741 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga
     1801 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac
     1861 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg
     1921 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcagtaat tctagagcgg
     1981 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2041 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2101 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2161 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2221 aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2341 tattgaaaaa ggaagaatcc tgaggccggg ccataacttc gtataatgta tgctatacga
     2401 agttatccat gggccccccc tcgacatgag taaacttggt ctgacagtta ccaatgctta
     2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3301 atactcatac tcttcctcag gttactcata tatactttag attgatttaa aacttcattt
     3361 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3421 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc gac
//
