LOCUS       sequence-482905-        6794 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  PiggyBac transposon encoding Neomycin resistance with deleted 
            inverted terminal repeat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  AUTHORS   Henssen AG, Henaff E, Jiang E, Eisenberg AR, Carson JR, Villasante 
            CM, Ray M, Still E, Burns M, Gandara J, Feschotte C, Mason CE, 
            Kentsis A
  TITLE     Genomic DNA transposition induced by human PGBD5.
  JOURNAL   Elife. 2015 Sep 25;4. pii: e10565. doi: 10.7554/eLife.10565.
  PUBMED    26406119
REFERENCE   2  (bases 1 to 6794)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-09-25"; volume: "4. pii: e10565. doi"; pages: " 
            10.7554/eLife.10565"
COMMENT     Sequence Label: PB-EF1-IRES-NEO deleted ITR
FEATURES             Location/Qualifiers
     source          1..6794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     712..734
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     726..743
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     727..743
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1321..1532
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1545..1813
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1883..2457,2458..2460,2461..2469)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2050..2067)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2277..2296
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2482..3276
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2536..2555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3146..3165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3341..3361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3403..3524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3440..3459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3494..3513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4773..4789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4773..4789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4786..4808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4797..4813
                     /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(4821..4827,4828..4845,4846..4851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4866..4887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5004..5021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5175..5763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5255..5274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5934..6725,6726..6794))
                     /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     6557..6576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtggggcctc ttcgctatta cgccagctgg
      661 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      721 gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg
      781 aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag atgaagatgc
      841 tcgacacgct gcagaacacg cagctagatg ttttatcggt ctgtatatcg aggtttattt
      901 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
      961 aacaaacaat ttatttatgt ttatttattt attaaaaaaa acaaaaactc aaaatttctt
     1021 ctataaagta acaaaacttt tatgagggac agcccccccc caaagccccc agggatgtaa
     1081 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     1141 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     1201 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     1261 gggatacggg gaaaaggcct ccacggccaa ggatctgcga tcgctccggt gcccgtcagt
     1321 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     1381 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     1441 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     1501 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     1561 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     1621 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     1681 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     1741 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     1801 gccgttacag atccaagctg tgaccggcgc ctactctaga gctagctcga gaattcgaat
     1861 ttaaatcgga tccgcggccg cgcccctctc cctccccccc ccctaacgtt actggccgaa
     1921 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1981 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     2041 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     2101 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     2161 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     2221 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     2281 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     2341 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     2401 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2461 gccacaacca tggcgtccgg aatgattgaa caagatggat tgcacgcagg ttctccggcc
     2521 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2581 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2641 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccgcgacg
     2701 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2761 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2821 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2881 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2941 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3001 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3061 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3121 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3181 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3241 atcgccttct atcgccttct tgacgagttc ttctgagtcg actgtacaat caacctctgg
     3301 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3361 gtggatacgc tgctttaatg cctttgtatc atgcgttaac taaacttgtt tattgcagct
     3421 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3481 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggaattga
     3541 ctcaaatgat gtcaattagt ctatcagaag ctcatctggt ctcccttccg ggggacaaga
     3601 catccctgtt taatatttaa acagcagtgt tcccaaactg ggttcttata tcccttgctc
     3661 tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta aagaaacagt
     3721 ggcagccagg tttagccccg gaattgactg gattcctttt ttagggccca ttggtatggc
     3781 tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag
     3841 gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg
     3901 ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag
     3961 cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc ctgatatcta
     4021 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     4081 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     4141 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     4201 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     4261 aaagagagag caatatttca agaatgtcta gctgcatcag gatcatatcg tcgggtcttt
     4321 tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg
     4381 tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg
     4441 cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca
     4501 cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt
     4561 ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga
     4621 cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta
     4681 cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga
     4741 gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4801 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4861 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4981 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5041 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5101 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5161 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5221 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5281 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5341 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5401 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5461 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5521 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5581 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5641 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5701 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5761 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5821 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5881 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5941 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6001 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6061 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6541 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6781 tgttgaatac tcat
//
