LOCUS       sequence-274027-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG sfGFP piggyBac transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7091)
  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            "Unpublished"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: PB CAG-sfGFP
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        147..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..805
                     /label=chicken beta-actin promoter
     intron          806..1822
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1746..1768
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1830..1849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1891..1893,1894..2607)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1891..2604
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1921..1949)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2544..2564
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    2644..3232
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2697..2717)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3115..3126)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3271..3288)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3277..3501
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3855..3889
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3976..3995)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3977..3995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4002..4019)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4002..4018)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4011..4033)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4160..4615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4297..4318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4509..4528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4642..4746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4747..4815,4816..5607)
                     /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(4965..4984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5778..6366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6267..6286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6520..6537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6654..6675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6690..6695,6696..6713,6714..6720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6728..6744
                     /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     6733..6755
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6752..6768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6752..6768
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6787..6807
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6789..6807
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   6871..6933
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 aagtaacaaa acttttaaac attctctctt ttacaaaaat aaacttattt tgtactttaa
       61 aaacagtcat gttgtattat aaaataagta attagcttaa cctatacata atagaaacaa
      121 attatactta ttaaccggtc atggtcgaca ttgattattg actagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccrgg
      661 gggggggggg gggggggggg gggggggggg ggsgrggcgg agaggtgcgg cggcagccaa
      721 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      781 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      841 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      901 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      961 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1021 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     1081 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     1141 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     1201 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     1261 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     1321 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     1381 gcaggtgggg gtgccgggcg gggcggggcc gcckcgggcc ggggrgggct cgggggrggg
     1441 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1501 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     1561 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1621 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1681 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1741 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1801 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     1861 ttttggcaaa gaattgctag cgccgccacc atgagcaaag gagaagaact tttcactgga
     1921 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
     1981 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
     2041 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
     2101 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
     2161 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
     2221 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
     2281 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
     2341 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
     2401 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
     2461 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
     2521 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
     2581 catggcatgg atgagctcta caaatgatta attaatgatc atacagtgct ctgagtacgc
     2641 tataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2701 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2761 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2821 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2881 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2941 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3001 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3061 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3121 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3181 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcaggatcca
     3241 ctagagcggc cgcctagagc tcgctatcag cctcgactgt gccttctagt tgccagccat
     3301 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3361 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3421 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3481 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag
     3541 cttgcggaac ccttaagctt ttttattata tattagtcac gatatctata acaagaaaat
     3601 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     3661 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     3721 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     3781 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     3841 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taaaaaagat
     3901 ttgcgcttta ctcgacctaa actttaaaca cgtcatagaa tcttcgtttg acaaaaacca
     3961 cattgtgttg ggtcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     4021 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4081 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4141 agcctgaatg gcgaatggca aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4201 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4261 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4321 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4381 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4441 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4501 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4561 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4621 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4681 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4741 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4801 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4861 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4921 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4981 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5041 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5101 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5161 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5221 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5281 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5341 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5401 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5461 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5521 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5581 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5641 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5701 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6421 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6481 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6541 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6601 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6661 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6721 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6781 aagcgcgcaa ttaaccctca ctaaagggaa cctcccccgg tattcacgac agcaggctga
     6841 ataataaaaa aattagaaac tattatttaa ccctagaaag ataatcatat tgtgacgtac
     6901 gttaaagata atcatgcgta aaattgacgc atgtgtttta tcggtctgta tatcgaggtt
     6961 tatttattaa tttgaataga tattaagttt tattatattt acacttacat actaataata
     7021 aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa
     7081 tttcttctat a
//
