LOCUS       sequence-486441-        7062 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  knock-in homology vector for ZF1 binding sites upstream of miniCMV 
            driving GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Fan V, Staller MV
  TITLE     Short activation domains control chromatin association of 
            transcription factors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pMVS254
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    396..1199
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1200..1225
                     /label=SA
                     /note="splice acceptor site"
     CDS             1249..1302
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1312..1710
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(1858..1875)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1864..1975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2316..2340
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2363..2372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3098..3163
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     misc_feature    3168..3756
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3221..3241)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3639..3650)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3770..3891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3781..3800)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3835..3854
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    3992..4828
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(4841..4857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4841..4857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4854..4876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4865..4881
                     /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(4889..4895,4896..4913,4914..4919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4934..4955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5072..5089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5243..5831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5323..5342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6002..6793,6794..6862))
                     /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     6625..6644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6863..6967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7035..7053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accagatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagttgctt tctctgacca gcattctctc
      421 ccctgggcct gtgccgcttt ctgtctgcag cttgtggcct gggtcacctc tacggctggc
      481 ccagatcctt ccctgccgcc tccttcaggt tccgtcttcc tccactccct cttccccttg
      541 ctctctgctg tgttgctgcc caaggatgct ctttccggag cacttccttc tcggcgctgc
      601 accacgtgat gtcctctgag cggatcctcc ccgtgtctgg gtcctctccg ggcatctctc
      661 ctccctcacc caaccccatg ccgtcttcac tcgctgggtt cccttttcct tctccttctg
      721 gggcctgtgc catctctcgt ttcttaggat ggccttctcc gacggatgtc tcccttgcgt
      781 cccgcctccc cttcttgtag gcctgcatca tcaccgtttt tctggacaac cccaaagtac
      841 cccgtctccc tggctttagc cacctctcca tcctcttgct ttctttgcct ggacaccccg
      901 ttctcctgtg gattcgggtc acctctcact cctttcattt gggcagctcc cctacccccc
      961 ttacctctct agtctgtgct agctcttcca gccccctgtc atggcatctt ccaggggtcc
     1021 gagagctcag ctagtcttct tcctccaacc cgggccccta tgtccacttc aggacagcat
     1081 gtttgctgcc tccagggatc ctgtgtcccc gagctgggac caccttatat tcccagggcc
     1141 ggttaatgtg gctctggttc tgggtacttt tatctgtccc ctccacccca cagtggggcc
     1201 tgacctcttc tcttcctccc acagggcctc gagagatctg gcagcggaga gggcagagga
     1261 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctaggctcga aatggccaag
     1321 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     1381 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     1441 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     1501 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     1561 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     1621 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     1681 ctgccctctg gttatgtgtg ggagggctaa cgcccgcccc acgacccgca gcgcccgacc
     1741 gaaaggagcg cacgacccca tggctccgac cgaagccacc cggggcggcc ccgccgaccc
     1801 cgcacccgcc cccgaggccc accgactcta gagggcccgt ttaaacccgc tgatcagcct
     1861 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1921 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1981 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2041 attgggaaga caatagcaac cggtcgggtt tcgtaacaat cgcatgagga ttcgcaacgc
     2101 cttcgtcgaa gtcgaagtcg acctcccgtc tcagtaaagg tcgtcgaagt cgaagtcgac
     2161 caatcggact gccttcgtac gtcgaagtcg aagtcgaccg tatcagtcgc ctcggaacgt
     2221 cgaagtcgaa gtcgaccatt cgtaagaggc tcactctccc ttacacggag tggataacta
     2281 gttaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
     2341 gcctggaacg cgtaccggtg tcgccaccat ggtgaagtac tacagtaaag gcgaagagct
     2401 ctttacaggt gtggtgccca tccttgtcga acttgatggg gacgtgaatg gccataaatt
     2461 ttctgtaagc ggtgaaggag agggcgatgc aacttacggt aagctcacac tcaagtttat
     2521 ctgcaccaca ggtaaactgc ccgtgccctg gccgaccctg gtcacgacgt tcggttacgg
     2581 tgtgcaatgc tttgcgcggt atccagatca tatgaaacaa catgactttt ttaagtccgc
     2641 gatgccggag gggtatgtcc aggaaaggac tatcttcttc aaggatgacg gtaactataa
     2701 gacacgggcc gaggttaaat tcgagggtga cacccttgtc aaccgcatcg aattgaaggg
     2761 gatagacttc aaggaagatg ggaatatact ggggcacaaa ctggaatata attataatag
     2821 tcacaacgtt tatattatgg cggataagca aaagaacgga atcaaggtga acttcaaaat
     2881 aaggcacaac atagaagacg gatccgtaca attggccgat cactatcaac aaaatacgcc
     2941 aataggggac ggcccagtcc ttctccccga caatcattat ctgtctactc aaagtgcgct
     3001 ctcaaaagac ccaaacgaga aaagagatca catggttctg ctggaatttg tgacagccgc
     3061 tggaatcaca cacggaatgg acgaacttta caaataagta aatataaaat ttttaagtgt
     3121 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagcgataat caacctctgg
     3181 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3241 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3301 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3361 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3421 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3481 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3541 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     3601 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3661 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3721 agacgagtcg gatctccctt tgggccgcct ccccgcatcg cagacatgat aagatacatt
     3781 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3841 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3901 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3961 taaaacctct acaaatgtgg tagcggccgc tactagggac aggattggtg acagaaaagc
     4021 cccatcctta ggcctcctcc ttcctagtct cctgatattg ggtctaaccc ccacctcctg
     4081 ttaggcagat tccttatctg gtgacacacc cccatttcct ggagccatct ctctccttgc
     4141 cagaacctct aaggtttgct tacgatggag ccagagagga tcctgggagg gagagcttgg
     4201 cagggggtgg gagggaaggg ggggatgcgt gacctgcccg gttctcagtg gccaccctgc
     4261 gctaccctct cccagaacct gagctgctct gacgcggctg tctggtgcgt ttcactgatc
     4321 ctggtgctgc agcttcctta cacttcccaa gaggagaagc agtttggaaa aacaaaatca
     4381 gaataagttg gtcctgagtt ctaactttgg ctcttcacct ttctagtccc caatttatat
     4441 tgttcctccg tgcgtcagtt ttacctgtga gataaggcca gtagccagcc ccgtcctggc
     4501 agggctgtgg tgaggagggg ggtgtccgtg tggaaaactc cctttgtgag aatggtgcgt
     4561 cctaggtgtt caccaggtcg tggccgcctc tactcccttt ctctttctcc atccttcttt
     4621 ccttaaagag tccccagtgc tatctgggac atattcctcc gcccagagca gggtcccgct
     4681 tccctaaggc cctgctctgg gcttctgggt ttgagtcctt ggcaagccca ggagaggcgc
     4741 tcaggcttcc ctgtccccct tcctcgtcca ccatctcatg cccctggctc tcctgcccct
     4801 tccctacagg ggttcctggc tctgctctgg cgtaatcatg gtcatagctg tttcctgtgt
     4861 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6121 ccagtgctgc aatgataccg cgactcccac gctcaccggc tccagattta tcagcaataa
     6181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7021 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
