LOCUS       sequence-486832-        8874 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  expression vector or PiggyBac-insertable cargo for expressing 
            HIF1a_R>D-SNAPtag-ERD-ZF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8874)
  AUTHORS   Fan V, Staller MV
  TITLE     Short activation domains control chromatin association of 
            transcription factors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8874)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pVF243
FEATURES             Location/Qualifiers
     source          1..8874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     481..500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(719..823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(849..1304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(986..1007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1198..1217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1431..1453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1445..1462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1446..1462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1592..1654
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2073..2572
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2108..2131)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2505..2524
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2594..3193
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2594..2613)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3090..3110
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3220..3237)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3226..3450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      3887..3896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5996..6037
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(6100..6120)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6162..6283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6199..6218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6253..6272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7046..7080
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7571..7587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7571..7587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7584..7606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7595..7611
                     /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(7619..7625,7626..7643,7644..7649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7664..7685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7802..7819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7973..8561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8053..8072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8732..8874,1..649,650..718))
                     /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"
ORIGIN
        1 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
       61 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      841 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
      901 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      961 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1021 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1081 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1141 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1201 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     1261 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     1321 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     1381 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     1441 acgttgtaaa acgacggcca gtgagcgcgc ctcgttcatt cacgtttttg aacccgtgga
     1501 ggacgggcag actcgcggtg caaatgtgtt ttacagcgtg atggagcaga tgaagatgct
     1561 cgacacgctg cagaacacgc agctagatta accctagaaa gataatcata ttgtgacgta
     1621 cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
     1681 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
     1741 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa
     1801 tttcttctat aaagtaacaa aacttttatg agggacagcc cccccccaaa gcccccaggg
     1861 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
     1921 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
     1981 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
     2041 cctgggggga tacggggaaa aggcgaacgc gtgggtaggg gaggcgcttt tcccaaggca
     2101 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     2161 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     2221 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     2281 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     2341 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     2401 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     2461 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     2521 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     2581 cccaagcttc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     2641 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     2701 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     2761 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2821 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     2881 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     2941 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     3001 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     3061 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     3121 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     3181 gcccggtgcc tgagcggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3241 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3301 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3361 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3421 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaacgcgat atgtacgggc
     3481 cagatatacg cgtcgcagaa caaacgccca gaaccgaagg ccggagccca gacgccgcag
     3541 cagcccagcc tgcgttgtaa cccggtcccc atagcaacca actaccgcag actactctcg
     3601 caagcgctgt tctgcttgtg acccggaccg gcttctcttc cgcttccggt cccacaatcc
     3661 tctgtcggcc tagaagagct ttgcattgtg ggagctcctt cctttctcgc tccccggcca
     3721 tcttggcggc tggtgttggt gagtgagctc tgcggggtaa acgattaggc ggctcgggga
     3781 gctccgctag ctggtgtttg acgctctgga ttataatcct ctcttctgtc ctctgtgtat
     3841 gctaggttgg gggttgtccc gcacctagag ctagcgccca gcttaagcca ccatggacct
     3901 gctgggccag agcatggacg agagcggcct gccccagctg accagctacg actgcgaggt
     3961 gaacgccccc atccagggca gcgacaacct gctgcagggc gaggagctgc tggacgccct
     4021 ggaccaggtg aacggatccg gcagcatgga caaagactgc gaaatgaagc gcaccaccta
     4081 cgatagccct ctgggcaagc tgctgctgtc tgggtgcgaa cagggcctgc accgtatcta
     4141 tttcgtgggc aatggacagg gagaacaagg ccctccagga ccagagccac tgatgcaggc
     4201 caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt tccctgtgcc
     4261 agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc tgtggaaact
     4321 gctgaaagtg gtgaagttcg gagaggtcat cagctacagc caactggccg ccctggccgg
     4381 caatcccgcc gccaccgccg ccgtgaaaac cgccctgaga ggaaatcccg tgcccattct
     4441 gatcccctgc caccgggtgg tgcagggcga cggcgacgtg gggggctacg agggccctct
     4501 ctacgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc ctgggctggg
     4561 tagcgctggc tctgctggag acatgagagc tgccaacctt tggccaagcc cgctcatgat
     4621 caaacgctct aagaagaaca gcctggcctt gtccctgacg gccgaccaga tggtcagtgc
     4681 cttgttggat gctgagcccc ccatactcta ttccgagtat gatcctacca gacccttcag
     4741 tgaagcttcg atgatgggct tactgaccaa cctggcagac agggagctgg ttcacatgat
     4801 caactgggcg aagagggtgc caggctttgt ggatttgacc ctccatgatc aggtccacct
     4861 tctagaatgt gcctggctag agatcctgat gattggtctc gtctggcgct ccatggagca
     4921 cccagtgaag ctactgtttg ctcctaactt gctcttggac aggaaccagg gaaaatgtgt
     4981 agagggcatg gtggagatct tcgacatgct gctggctaca tcatctcggt tccgcatgat
     5041 gaatctgcag ggagaggagt ttgtgtgcct caaatctatt attttgctta attctggagt
     5101 gtacacattt ctgtccagca ccctgaagtc tctggaagag aaggaccata tccaccgagt
     5161 cctggacaag atcacagaca ctttgatcca cctgatggcc aaggcaggcc tgaccctgca
     5221 gcagcagcac cagcggctgg cccagctcct cctcatcctc tcccacatca ggcacatgag
     5281 taacaaaggc atggagcatc tgtacagcat gaagtgcaag aacgtggtgc ccctctatga
     5341 cctgctgctg gagatgctgg acgcccaccg cctacatgcg cccactagcc gtggaggggc
     5401 atccgtggag gagacggacc aaagccactt ggccactgcg ggctctactt catcgggtac
     5461 ctctagacca ggcgaacgac cgtttcaatg ccggatatgt atgaggaact tctccgagga
     5521 ggcaaacttg aggcgccaca cccgaacaca tacaggagaa aagccattcc aatgtcgaat
     5581 ttgtatgcgc aatttttcag atcactcaag cctcaagcga cacctccgca cacatactgg
     5641 ttcacagaag ccctttcagt gcaggatttg tatgcgcaac tttagccaat cagcgaacct
     5701 tttgcggcac actagaacgc atacaggtga gaagcctttc cagtgtcgca tctgtatgcg
     5761 gaacttcagc gaccccagtt cattgaagag gcatttgcga actcacaccg gttctcaaaa
     5821 accttttcag tgccgaattt gtatgcgcaa cttcagccaa caaacaaatt tgacgagaca
     5881 cacgcgcacg cacaccgggg aaaaaccgtt tcaatgtcga atctgcatgc gcaattttag
     5941 cgatgcgaca caacttgtta ggcatctgcg cacacacttg cggggaagcg gagctggtaa
     6001 gcctatccct aaccctctcc tcggtctcga ttctacctga gtcgacaatc aacctctgga
     6061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6121 tggatacgct gctttaatgc ctttgtatca tgcgttaact aaacttgttt attgcagctt
     6181 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6241 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggaattgac
     6301 tcaaatgatg tcaattagtc tatcagaagc tcatctggtc tcccttccgg gggacaagac
     6361 atccctgttt aatatttaaa cagcagtgtt cccaaactgg gttcttatat cccttgctct
     6421 ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg aagtccctaa agaaacagtg
     6481 gcagccaggt ttagccccgg aattgactgg attccttttt tagggcccat tggtatggct
     6541 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     6601 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     6661 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     6721 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc tgatatctat
     6781 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
     6841 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
     6901 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
     6961 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
     7021 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
     7081 ttaatctagc tgcatcagga tcatatcgtc gggtcttttt tccggctcag tcatcgccca
     7141 agctggcgct atctgggcat cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa
     7201 gcggcatgga aagagtttgc cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc
     7261 acgtttacca tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc
     7321 gttcaggcca cctgggatac cagttcgtcg cggcttttcc ggacacagtt ccggatggtc
     7381 agcccgaagc gcatcagcaa cccgaacaat accggcgaca gccggaactg ccgtgccggt
     7441 gtgcagatta atgacagcgg tgcggcgctg ggatattacg tcagcgagga cgggtatcct
     7501 ggctggatgc cgcagaaatg gacatggata ccccgtgagt tacccggcgg gcgcgcttgg
     7561 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7621 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7681 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7741 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7801 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8761 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8821 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgag
//
