LOCUS       sequence-258678-        5036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized Sleeping Beauty Transposon Vector for GFP expression by a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5036)
  TITLE     Optimized Sleeping Beauty Transposon Vector with CMV-GFP cassette
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5036)
  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            "Unpublished"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: pT4-CMV-GFP
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          676..808
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      813..822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(819..821,822..824,825..1538)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(819..821,822..824,825..1535)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(864..885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1125..1144)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1472..1493
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1545..1593
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1929..1949)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1929..1947)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1968..1984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1968..1984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1981..2003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1992..2008
                     /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(2016..2022,2023..2040,2041..2046))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2061..2082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2199..2216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2370..2958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2450..2469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3129..3920,3921..3989))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3752..3771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3990..4094)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4121..4576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4208..4227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4418..4439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4702..4724
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4716..4733
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4717..4733
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4743..4762
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4743..4761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aggtcgttta gtgaaccgtc
      601 agatcactag tagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      661 ctcgactgat cacaggtaag tatcaaggtt acaagacagg tttaaggagg ccaatagaaa
      721 ctgggcttgt cgagacagag aagattcttg cgtttctgat aggcacctat tggtcttact
      781 gacatccact ttgcctttct ctccacaggg cggccgccat ggtgagcaag ggcgaggagc
      841 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      901 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      961 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1021 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1081 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1141 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1201 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1261 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1321 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1381 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1441 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1501 ccgggatcac tctcggcatg gacgagctgt acaagtaaga attcaataaa atatctttat
     1561 tttcattaca tctgtgtgtt ggttttttgt gtgtaattcg agcatcttac cgccatttat
     1621 tcccagatct ttaaacaatt taaaggcaat gctaccaaat actaagcgcg tgtatgtaca
     1681 cttctgaccc actgggaatg tgatgaaaga aataaaagct gaaatgaatc attctctcta
     1741 ctattattct gatatttcac attcttaaaa taaagtggtg atcctaactg accttaagac
     1801 agggaatctt tactcggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt
     1861 ggctaaggtg tatgtaaact tccgacttca actgtagttt aaacgggccc ggtacccagc
     1921 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     1981 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2041 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2161 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2221 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2281 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2341 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2401 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2461 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2521 tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc acgctgtagg
     2581 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2641 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2701 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2761 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2821 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2881 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2941 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3001 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3061 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3121 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3181 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3241 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3301 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3361 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3421 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3481 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3541 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt ggccgcagtg
     3601 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3661 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3721 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3781 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3841 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3901 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3961 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4021 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4081 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     4141 ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4321 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4441 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4561 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4621 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     4681 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     4741 cgtaatacga ctcactatag ggcgaattgg agctagcgtt taaacctaca gttgaagtcg
     4801 gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta ctccacaaat
     4861 ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt gtgcatgaca
     4921 caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat tcactgtatc
     4981 acaattccag tgggtcagaa gtgtacatac acgcgcttga ctgtgccttt aagctt
//
