LOCUS       sequence-420275-        8484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac vector to express MLV Gag with truncated MA domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8484)
  AUTHORS   Najia MA, Borrajo J, Le A, Tsai F, Huang JY, Griffith LG, Daley GQ, 
            Blainey PC
  TITLE     Live-cell transcriptomics with engineered virus-like particles
  JOURNAL   bioRxiv 2024.10.01.616098
REFERENCE   2  (bases 1 to 8484)
  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: "bioRxiv 
            2024.10.01.616098"
COMMENT     Sequence Label: pPB_EF1a_Gag∆MA
FEATURES             Location/Qualifiers
     source          1..8484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   127..189
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1356..2534
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1587..2525
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2482..2502
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3504..3560
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      3810..3819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4307..4428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4344..4363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4398..4417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5191..5225
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5716..5732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5716..5732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5729..5751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5740..5756
                     /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(5764..5770,5771..5788,5789..5794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5809..5830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5947..5964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6118..6706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6198..6217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6877..7668,7669..7737))
                     /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     7500..7519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7738..7842)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7868..8323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8005..8026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8217..8236
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8450..8472
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8464..8481
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8465..8481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg ggcagactcg cggtgcaaat
       61 gtgttttaca gcgtgatgga gcagatgaag atgctcgaca cgctgcagaa cacgcagcta
      121 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgtgtaaaat
      181 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt
      241 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg
      301 tttatttatt tattaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaactt
      361 ttatgaggga cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta
      421 gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg
      481 agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc
      541 tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaaggcc
      601 tccaccataa agatggttaa ttaacccacc caagatctgg atcgataagc ttgatatcga
      661 attcgggagg tggtccctgc agttacgcca atgataaccc ccgccagaaa aatcttagta
      721 gccttccctt tttgttttcc gtgccccaac tcggcggatt gactcggccc cttccggaaa
      781 cacccgaatc aacttctagt caaattattg ttcacgccgc aatgacccac ccctggcccg
      841 cgtctgtgga actgacccct ggtgtacagg agagttcgct gctgaaagtg gtcccaaagg
      901 ggtactagtt tttaagctcc caactccccc tcccccagcg tctggaggat tccacaccct
      961 cgcaccgcag gggcgaggaa gtgggcggag tccggttttg gcgccagccg ctgaggctgc
     1021 caagcagaaa agccaccgct gaggagactc cggtcactgt cctcgccccg cctccccctt
     1081 ccctcccctt ggggaccacc gggcgccacg ccgcgaacgg taagtgccgc ggtcgtcggc
     1141 gcctccgccc tccccctagg gccccaattc ccagcgggcg cggcgcgcgg ccccgcgcgc
     1201 agctcccggc tccctccccc ttcggatgtg gcttgagctg taggcgcgga gtcctgcagc
     1261 cccgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     1321 ctgtaggttt ggcaatcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     1381 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg
     1441 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     1501 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1561 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     1621 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     1681 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     1741 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     1801 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     1861 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     1921 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     1981 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     2041 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     2101 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     2161 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     2221 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     2281 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     2341 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     2401 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     2461 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     2521 ttcaggtgtc gtgacgtacg gccaccatgc ccctccgcgc aggaggaaac ggacagcttc
     2581 aatactggcc gttctcctct tctgaccttt acaactggaa aaataataac ccttcttttt
     2641 ctgaagatcc aggtaaactg acagctctga tcgagtctgt cctcatcacc catcagccca
     2701 cctgggacga ctgtcagcag ctgttgggga ctctgctgac cggagaagaa aaacaacggg
     2761 tgctcttaga ggctagaaag gcggtgcggg gcgatgatgg gcgccccact caactgccca
     2821 atgaagtcga tgccgctttt cccctcgagc gcccagactg ggattacacc acccaggcag
     2881 gtaggaacca cctagtccac tatcgccagt tgctcctagc gggtctccaa aacgcgggca
     2941 gaagccccac caatttggcc aaggtaaaag gaataacaca agggcccaat gagtctccct
     3001 cggccttcct agagagactt aaggaagcct atcgcaggta cactccttat gaccctgagg
     3061 acccagggca agaaactaat gtgtctatgt ctttcatttg gcagtctgcc ccagacattg
     3121 ggagaaagtt agagaggtta gaagatttaa aaaacaagac gcttggagat ttggttagag
     3181 aggcagaaaa gatctttaat aaacgagaaa ccccggaaga aagagaggaa cgtatcagga
     3241 gagaaacaga ggaaaaagaa gaacgccgta ggacagagga tgagcagaaa gagaaagaaa
     3301 gagatcgtag gagacataga gagatgagca agctattggc cactgtcgtt agtggacaga
     3361 aacaggatag acagggagga gaacgaagga ggtcccaact cgatcgcgac cagtgtgcct
     3421 actgcaaaga aaaggggcac tgggctaaag attgtcccaa gaaaccacga ggacctcggg
     3481 gaccaagacc gcagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     3541 tcgaagagaa tcctggaccg gtcagcaagg gcgaggagga caacgcctct ctcccagcga
     3601 cacacgagtt acacatcttt ggctccatca acggtgtgga cttcgacctc gtgggtcagg
     3661 gcaccggcaa tccaaacgac ggttacgagg agctcaacct caagtccacc aagggcgacc
     3721 tccagttctc cccctggatt ctggtccctc atatcgggta cggcttccat cagtacctgc
     3781 cctacccaga cgggatgtcg cctttccagg ccgccatggt ggacggctcc ggataccaag
     3841 tccatcgcac aatgcagttc gaagacggtg cctcccttac tgtcaactac cgctacacct
     3901 acgagggaag ccacatcaaa ggagaggccc aggtcaaggg gactggtttc cctgccgacg
     3961 gtcctgtcct caccaactcg ctcaccgctg cggactggtg caggtcgaag aagacttacc
     4021 ccaacgacaa aaccatcatc agtaccttca agtggagtta caccactgga aacggcaagc
     4081 gctaccggag cactgcgcgg accacctaca cctttgccaa gccaatggcg gccaactatc
     4141 tcaagaacca gccgatgtac gtgttccgca agacggagct caagcactcc aagaccgagc
     4201 tcaacttcaa ggagtggcaa aaggccttta ccgacgtcat gggcatggac gagctctaca
     4261 agtaacaatc aacctctgga ttacaaaatt tgtggttaac taactaaact tgtttattgc
     4321 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4381 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggaa
     4441 ttgactcaaa tgatgtcaat tagtctatca gaagctcatc tggtctccct tccgggggac
     4501 aagacatccc tgtttaatat ttaaacagca gtgttcccaa actgggttct tatatccctt
     4561 gctctggtca accaggttgc agggtttcct gtcctcacag gaacgaagtc cctaaagaaa
     4621 cagtggcagc caggtttagc cccggaattg actggattcc ttttttaggg cccattggta
     4681 tggctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc
     4741 agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc
     4801 tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc
     4861 ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct gtccctgata
     4921 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     4981 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     5041 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     5101 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     5161 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     5221 tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg ctcagtcatc
     5281 gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg
     5341 ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac gttgagcgaa
     5401 aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt aacggtgaac
     5461 tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca cagttccgga
     5521 tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg aactgccgtg
     5581 ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc gaggacgggt
     5641 atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc ggcgggcgcg
     5701 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5761 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5821 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6541 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6661 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6721 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6781 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6841 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7081 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7141 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7741 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7861 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     7921 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     7981 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     8041 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     8101 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     8161 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     8221 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     8281 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     8341 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8401 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8461 cgttgtaaaa cgacggccag tgag
//
