LOCUS       sequence-420658-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac donor vector containing LacI-eGFP for random integration by
            PBase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  AUTHORS   Mach P, Kos PI, Zhan Y, Cramard J, Gaudin S, Tunnermann J, Marchi E,
            Eglinger J, Zuin J, Kryzhanovska M, Smallwood S, Gelman L, Roth G, 
            Nora EP, Tiana G, Giorgetti L
  TITLE     Cohesin and CTCF control the dynamics of chromosome folding.
  JOURNAL   Nat Genet. 2022 Dec;54(12):1907-1918. doi: 
            10.1038/s41588-022-01232-7. Epub 2022 Dec 5.
  PUBMED    36471076
REFERENCE   2  (bases 1 to 8173)
  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"; doi: 
            "10.1038/s41588-022-01232-7"; journalName: "Nat Genet"; date: 
            "2022-12"; volume: "54"; issue: "12"; pages: "1907-1918"
COMMENT     Sequence Label: PB-LacI-eGFP
FEATURES             Location/Qualifiers
     source          1..8173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2237..2271
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2552..2921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4862..4881
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             6021..6041
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      6132..6141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6138..6140,6141..6143,6144..6857)
                     /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(6138..6140,6141..6143,6144..6854)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6183..6204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6444..6463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6791..6812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6918..6942)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7125..7246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7162..7181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7216..7235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7432..7494
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7529..7548)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7530..7548)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7558..7575)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7558..7574)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7567..7589)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7716..8171
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7803..7822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8013..8034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggggcgc
     2221 gccgcagcta gattaaccct agaaagatag tctgcgtaaa attgacgcat gcattcttga
     2281 aatattgctc tctctttcta aatagcgcga atccgtcgct gtgcatttag gacatctcag
     2341 tcgccgcttg gagctcccgt gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga
     2401 actataacaa ccgcgtgagt caaaatgacg catgattatc ttttacgtga cttttaagat
     2461 ttaactcata cgataattat attgttattt cgtgttctac ttacgtgata acttattata
     2521 tatatatttt cttgttatag atatccttct cgacattgat tattgactag agatctatca
     2581 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2641 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2701 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2761 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2821 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2881 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
     2941 tctgcttcac tctccccatc tccccccccc tccccacccc caattttgta tttatttatt
     3001 ttttaattat tttgtgcagc gatgggggcg aggggcgggg cggggcgagg cggagaggtg
     3061 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     3121 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
     3181 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     3241 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
     3301 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     3361 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     3421 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     3481 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     3541 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     3601 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     3661 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     3721 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     3781 gctcggggga gggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     3841 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     3901 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     3961 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     4021 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     4081 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctg tgagcgctgc
     4141 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     4201 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     4261 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     4321 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     4381 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     4441 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     4501 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     4561 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     4621 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     4681 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     4741 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     4801 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     4861 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgagcctgac
     4921 aggtgtgaaa caggaagaga accggtacca tggtaaaata cgtgactttg tacgatgtag
     4981 ctgaatacgc aggagtgagc tatcagaccg tctcacgagt agtcaatcag gccagtcatg
     5041 tctcagcaaa gaccagggag aaagtagaag ccgcaatggc cgagctgaat tacattccta
     5101 acagagtagc ccagcaactt gcagacaagg gtggcggttc cctcttgata ggagtagcct
     5161 catcctcact ggccttgcat gcaccatccc agatagtagc tgctattaaa tctcgcgctg
     5221 atcagttggg ggcctcagtg gtagtttcta tggtcgagag gtccggtgtg gaagcctgta
     5281 aagccgccgt tcacaatctc ctcgcacaac gcgtctctgg attgattatt aactaccctt
     5341 tggatgacca ggacgctata gctgtggaag ctgcctgcac caatgttcct gccttgttct
     5401 tggatgtgag tgatcaaact ccaataaatt ccataatttt ttcacatgaa gatggaacaa
     5461 gacttggagt ggaacacctg gtcgctctgg gccatcaaca gatagcattg ctcgccgggc
     5521 cacttagtag cgtcagtgca cggctgcgct tggccgggtg gcacaaatac ctgaccagga
     5581 accaaatcca accaattgca gagagagagg gagattggag tgcaatgagc ggatttcaac
     5641 aaacaatgct tatgcttaat gaaggcatag ttcccaccgc aatgctggtt gcaaacgatc
     5701 aaatggccct gggggcaatg cgcgccataa cagagtctgg cttgcgagtc ggagccgata
     5761 taagcgtcgt gggatatgac gatacagaag acagcagctg ttacattcct cctctcacaa
     5821 ccatcaaaca ggactttaga ctgctcggac aaacatcagt ggaccgcctg ctgcaacttt
     5881 ctcaaggcca agctgtcaaa gggaatcagc tcctgcctgt tagtctcgtt aagcgaaaaa
     5941 caaccttggc cccaaacaca caaactgctt cccctagggc tctggctgat tcattgatgc
     6001 aattgggctc atgtacaccc cctaaaaaaa agcgaaaggt agccgcggat ctaaaggcac
     6061 gtcgtccagc actaagatca ccaccactag gaactcgtaa gcgaaagcgt ccttctcggg
     6121 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6181 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     6241 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6301 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     6361 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6421 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     6481 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6541 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     6601 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     6661 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     6721 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     6781 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     6841 acgagctgta caagtaaagg aattctggcc atctagaagc tcgctttctt gctgtccaat
     6901 ttcctcgaga tatctagacc cagctttctt gtacaaagtg gtgatcctta tttattgatc
     6961 cttatttatt gtagcttatt tattgctact tatttattga tctgcggccg cgactctaga
     7021 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     7081 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     7141 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     7201 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaagga attcgataaa
     7261 agttttgtta ctttatagaa gaaattttga gtttttgttt ttttttaata aataaataaa
     7321 cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt
     7381 aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca
     7441 attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct agggttaatc
     7501 tagctgcggc gcgccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     7561 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     7621 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     7681 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     7741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     8041 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     8101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     8161 gcttacaatt tag
//
