LOCUS       sequence-355024-        4973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon vector prepared for Esp3I golden gate cloning of 
            Transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4973)
  AUTHORS   Pallares-Masmitja M, Ivancic D, Mir-Pedrol J, Jaraba-Wallace J, 
            Tagliani T, Oliva B, Rahmeh A, Sanchez-Mejias A, Guell M
  TITLE     Find and cut-and-transfer (FiCAT) mammalian genome engineering.
  JOURNAL   Nat Commun. 2021 Dec 3;12(1):7071. doi: 10.1038/s41467-021-27183-x.
  PUBMED    34862378
REFERENCE   2  (bases 1 to 4973)
  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: "Nat 
            Commun."; date: "2021-12-3"; volume: "12(1):7071. doi"; pages: " 
            10.1038/s41467-021-27183-x"
COMMENT     Sequence Label: pTSB-FICAT_Payload-Esp3I
FEATURES             Location/Qualifiers
     source          1..4973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(409..1200,1201..1269))
                     /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     1032..1051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1270..1374)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1400..1855)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1537..1558)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1749..1768
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1982..2004
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1996..2013
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1997..2013
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2143..2205
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             join(2643..2645,2646..2648,2649..3161)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(2688..2709)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2949..2968)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     repeat_region   3696..3730
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4221..4237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4221..4237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4234..4256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4245..4261
                     /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(4269..4275,4276..4293,4294..4299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4314..4335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4452..4469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4623..4973,1..238))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4703..4722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
       61 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      121 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      181 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      241 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      301 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      541 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      601 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1381 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1441 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1501 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1561 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1621 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1681 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1741 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1801 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     1861 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1921 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1981 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt
     2041 gaacccgtgg aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag
     2101 atgaagatgc tcgacacgct gcagaacacg cagctagatt aaccctagaa agataatcat
     2161 attgtgacgt acgttaaaga taatcatgtg taaaattgac gcatgtgttt tatcggtctg
     2221 tatatcgagg tttatttatt aatttgaata gatattaagt tttattatat ttacacttac
     2281 atactaataa taaattcaac aaacaattta tttatgttta tttatttatt aaaaaaaaca
     2341 aaaactcaaa atttcttcta taaagtaaca aaacttttat gagggacagc ccccccccaa
     2401 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
     2461 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
     2521 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
     2581 gcctgcagac acctgggggg atacggggaa aaggcctcct cggccactag ttatgcgaga
     2641 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2761 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     2821 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3061 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     3121 acggcatcaa ggccaacttc aagatccgcc acaacatcga gcgtctccta acgggcccat
     3181 tggtatggct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag
     3241 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg
     3301 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct
     3361 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc
     3421 tgatatctat aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat
     3481 aacaatataa ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc
     3541 attttgactc acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca
     3601 cgcctcacgg gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc
     3661 gctatttaga aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat
     3721 ctttctaggg ttaatctagc tgcatcagga tcatatcgtc gggtcttttt tccggctcag
     3781 tcatcgccca agctggcgct atctgggcat cggggaggaa gaagcccgtg ccttttcccg
     3841 cgaggttgaa gcggcatgga aagagtttgc cgaggatgac tgctgctgca ttgacgttga
     3901 gcgaaaacgc acgtttacca tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg
     3961 tgaactgttc gttcaggcca cctgggatac cagttcgtcg cggcttttcc ggacacagtt
     4021 ccggatggtc agcccgaagc gcatcagcaa cccgaacaat accggcgaca gccggaactg
     4081 ccgtgccggt gtgcagatta atgacagcgg tgcggcgctg ggatattacg tcagcgagga
     4141 cgggtatcct ggctggatgc cgcagaaatg gacatggata ccccgtgagt tacccggcgg
     4201 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agc
//
