LOCUS       sequence-486836-        8736 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  expression vector or PiggyBac-insertable cargo for expressing 
            noAD-SNAPtag-ERD-ZF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8736)
  AUTHORS   Fan V, Staller MV
  TITLE     Short activation domains control chromatin association of 
            transcription factors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8736)
  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            "Unpublished"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: pVF230
FEATURES             Location/Qualifiers
     source          1..8736
                     /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             5858..5899
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5962..5982)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6024..6145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6061..6080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6115..6134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6908..6942
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7433..7449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7433..7449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7446..7468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7457..7473
                     /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(7481..7487,7488..7505,7506..7511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7526..7547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7664..7681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7835..8423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7915..7934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8594..8736,1..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 ccatgggatc
     3901 cggcagcatg gacaaagact gcgaaatgaa gcgcaccacc tacgatagcc ctctgggcaa
     3961 gctgctgctg tctgggtgcg aacagggcct gcaccgtatc tatttcgtgg gcaatggaca
     4021 gggagaacaa ggccctccag gaccagagcc actgatgcag gccaccgcct ggctcaacgc
     4081 ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc accacccagt
     4141 gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag tggtgaagtt
     4201 cggagaggtc atcagctaca gccaactggc cgccctggcc ggcaatcccg ccgccaccgc
     4261 cgccgtgaaa accgccctga gaggaaatcc cgtgcccatt ctgatcccct gccaccgggt
     4321 ggtgcagggc gacggcgacg tggggggcta cgagggccct ctctacgtga aagagtggct
     4381 gctggcccac gagggccaca gactgggcaa gcctgggctg ggtagcgctg gctctgctgg
     4441 agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa
     4501 cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc
     4561 ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg
     4621 cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt
     4681 gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct
     4741 agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga agctactgtt
     4801 tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat
     4861 cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga
     4921 gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag
     4981 caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga
     5041 cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct
     5101 ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca
     5161 tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc tggagatgct
     5221 ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacgga
     5281 ccaaagccac ttggccactg cgggctctac ttcatcgggt acctctagac caggcgaacg
     5341 accgtttcaa tgccggatat gtatgaggaa cttctccgag gaggcaaact tgaggcgcca
     5401 cacccgaaca catacaggag aaaagccatt ccaatgtcga atttgtatgc gcaatttttc
     5461 agatcactca agcctcaagc gacacctccg cacacatact ggttcacaga agccctttca
     5521 gtgcaggatt tgtatgcgca actttagcca atcagcgaac cttttgcggc acactagaac
     5581 gcatacaggt gagaagcctt tccagtgtcg catctgtatg cggaacttca gcgaccccag
     5641 ttcattgaag aggcatttgc gaactcacac cggttctcaa aaaccttttc agtgccgaat
     5701 ttgtatgcgc aacttcagcc aacaaacaaa tttgacgaga cacacgcgca cgcacaccgg
     5761 ggaaaaaccg tttcaatgtc gaatctgcat gcgcaatttt agcgatgcga cacaacttgt
     5821 taggcatctg cgcacacact tgcggggaag cggagctggt aagcctatcc ctaaccctct
     5881 cctcggtctc gattctacct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5941 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6001 gcctttgtat catgcgttaa ctaaacttgt ttattgcagc ttataatggt tacaaataaa
     6061 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6121 tgtccaaact catcaatgta tcttatcatg tctggaattg actcaaatga tgtcaattag
     6181 tctatcagaa gctcatctgg tctcccttcc gggggacaag acatccctgt ttaatattta
     6241 aacagcagtg ttcccaaact gggttcttat atcccttgct ctggtcaacc aggttgcagg
     6301 gtttcctgtc ctcacaggaa cgaagtccct aaagaaacag tggcagccag gtttagcccc
     6361 ggaattgact ggattccttt tttagggccc attggtatgg ctttttcccc gtatcccccc
     6421 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     6481 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     6541 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     6601 catccctggg ggctttgggg gggggctgtc cctgatatct ataacaagaa aatatatata
     6661 taataagtta tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat
     6721 cttaaaagtc acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc
     6781 aaaatcagtg acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact
     6841 gagatgtcct aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc
     6901 aagaatgcat gcgtcaattt tacgcagact atctttctag ggttaatcta gctgcatcag
     6961 gatcatatcg tcgggtcttt tttccggctc agtcatcgcc caagctggcg ctatctgggc
     7021 atcggggagg aagaagcccg tgccttttcc cgcgaggttg aagcggcatg gaaagagttt
     7081 gccgaggatg actgctgctg cattgacgtt gagcgaaaac gcacgtttac catgatgatt
     7141 cgggaaggtg tggccatgca cgcctttaac ggtgaactgt tcgttcaggc cacctgggat
     7201 accagttcgt cgcggctttt ccggacacag ttccggatgg tcagcccgaa gcgcatcagc
     7261 aacccgaaca ataccggcga cagccggaac tgccgtgccg gtgtgcagat taatgacagc
     7321 ggtgcggcgc tgggatatta cgtcagcgag gacgggtatc ctggctggat gccgcagaaa
     7381 tggacatgga taccccgtga gttacccggc gggcgcgctt ggcgtaatca tggtcatagc
     7441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8701 taccatctgg ccccagtgct gcaatgatac cgcgag
//
