LOCUS       sequence-101284-        3750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A directional cloning vector of PCR-amplified cDNAs for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3750)
  TITLE     Directional cloning vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3750)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pUC-dirTopo-AU1-aGFP
FEATURES             Location/Qualifiers
     source          1..3750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    637..653
                     /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)."
     CDS             join(898..900,901..1614)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(898..900,901..1611)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(940..961)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1201..1220)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1548..1569
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1741..1862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1778..1797)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1832..1851
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1912..2016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2017..2085,2086..2877)
                     /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(2235..2254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3048..3636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3537..3556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagctt
      601 tacactttat tcttccggct cgtattttgt gtggaattgt gagcggataa caatttccgg
      661 aggaggccct tcacctgaag agcggacgga cgagcggata tcgccgccta accccgctct
      721 tcaaagggat ccaatgacac ctacaggtac atcgtcgaca ttaccgattc actggccgtc
      781 gttctgcaac gccgcgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      841 catccccctt tcgccagttg gcgcaatagc gaagaggccc gcaccgaccg cgaattcgtg
      901 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      961 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1021 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1081 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1141 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1201 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1261 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1321 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1381 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1441 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1501 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1561 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaactcgag
     1621 taatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1681 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1741 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1801 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1861 taacgcgtaa attgtaagcg ttaatcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1921 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1981 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2041 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2101 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2161 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2221 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2281 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2341 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2401 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2461 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2521 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2581 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2641 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2701 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2761 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2821 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2881 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2941 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3001 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3061 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3181 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3721 tctgtggata accgtattac cgccatgcat
//
