LOCUS       sequence-223679-        5950 bp    DNA     circular SYN 14-APR-2025
DEFINITION  evaluate FOXA1 DIV motif enhancer activity near 50 kb of ATP9A gene 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5950)
  AUTHORS   Wang X, Srivastava Y, Jankowski A, Malik V, Wei Y, Del Rosario RC, 
            Cojocaru V, Prabhakar S, Jauch R
  TITLE     DNA-mediated dimerization on a compact sequence signature controls 
            enhancer engagement and regulation by FOXA1.
  JOURNAL   Nucleic Acids Res. 2018 Jun 20;46(11):5470-5486. doi: 
            10.1093/nar/gky259.
  PUBMED    29669022
REFERENCE   2  (bases 1 to 5950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2018-06-20- 20"; volume: "46"; issue: 
            "11"; pages: "5470-5486"
COMMENT     Sequence Label: ATP9A-Tol2
FEATURES             Location/Qualifiers
     source          1..5950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(690..709)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(691..709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(716..733)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(716..732)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(725..747)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      873..1328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(960..979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1170..1191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1406..1510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1511..1579,1580..2371)
                     /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     complement(1729..1748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2542..3130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3031..3050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3284..3301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3418..3439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3454..3459,3460..3477,3478..3484)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3492..3508
                     /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)."
     primer_bind     3497..3519
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3516..3532
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3516..3532
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3550..3568
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3550..3567
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(4908..4910,4911..4913,4914..5627)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4908..4910,4911..4913,4914..5624)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4953..4974)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5214..5233)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5561..5582
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5750..5871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5787..5806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5841..5860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 aactatcagt aaaacaaaat acttaatgta cttacctcaa tatgcttcct taggtttgat
       61 ggcgaacttt tgaaggccga tatttcttta ttaagcggga gacagaggac acatttcatc
      121 ttgaatgaat ctttatttac accacttaaa gaaaagaatt cgcgaagata cggccacggg
      181 tgctcttgat cctgtggctg attttggact gtgctgctcg cagctgctga tgaatcacat
      241 acttcctcca ttttcttcca ctgattgact gttataattt ccctaatttc caggtcaagg
      301 tgctgtgcat tgtggtaata gatgtgacat gacgtcactt ccaaaggacc aatgaacatg
      361 tctgaccaat ttcatataat gtgaaaacga ttttcatagg cagaataaat aacatttaaa
      421 ttaaactggg catcagcgca attcaattgg tttggtaata gcaagggaaa atagaatgaa
      481 gtgatctcca aaaaataagt actttttgac tgtaaataaa attgtaagga gtaaaaagta
      541 cttttttttc taaaaaaatg taattaagta aaagtaaaag tattgatttt taattgtact
      601 caagtaaagt aaaaatcccc aaaaataata cttaagtaca gtaatcaagt aaaattactc
      661 aagtacttta cacctctggg cccaattcgc cctatagtga gtcgtattac aattcactgg
      721 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      781 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      841 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
      901 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      961 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1021 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1081 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1141 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1201 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1261 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     1321 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc
     1381 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1441 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1501 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2041 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2221 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2341 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2461 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3181 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3241 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3301 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3361 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3421 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3481 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3541 cgccaagcta tttaggtgac actatagaat actcaagcta tgcatccaac gcgttgggag
     3601 ctctcccata tggtcgagca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
     3661 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
     3721 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa
     3781 gtaaaagcaa gaaagaaaac tagagattct tgtttaagct tttaatctca aaaaacatta
     3841 aatgaaatgc atacaaggtt ttatcctgct ttagaactgt ttgtatttaa ttatcaaact
     3901 ataagacaga caatctaatg ccagtacacg ctactcaaag ttgtaaaacc tcagatttaa
     3961 cttcagtaga agctgattct caaaattgtt agtgtcaagc ctagctcttt tggggctgaa
     4021 aagcaatcct gcagtcgacg tcggccatag cggccgcgga agcctcagtc ttccaaagtg
     4081 gattacaagc atgagccacc gcgcccagcc tagaccattt atataaagga aaaccaaaat
     4141 tgattttctt aggatttgac tttaagacaa ataaaggttt taacagcaga tagtctgcta
     4201 accagatgtg caatggtttc atacaatctg gctgaaatgc aaaagcaaat attagaggag
     4261 cagagccaag ggaacaatgt ttctgaaaaa ggtaactaat gtaacaagga gttggcctcc
     4321 acagaaacca tcatctctgc tgggagttcc cctataatca gttagcactg cgcaaatatt
     4381 tgctgaattg tctgcatgac acaccaacct tccaggaagt tacagggtcc aggaaagaac
     4441 agtctcagca ttgacgctgg gcagatccaa gaagaacatg aagaaaaatt aaaagtagcc
     4501 agatatggtg gctcatgcct gtaatcccag tactttggga ggccgaggcg ggagaatcac
     4561 ttgaggtcag gagttcgaga ccagcctggc caacatgacg aaaccttgtc tctactaaaa
     4621 atataaaaat tagctgggca tgggccgggt gcagtggctg actcctgtaa tcccagcact
     4681 ttgggaggcc aaggcgggtg gatcaagagg tcaggagatc aagaccattc tggccaacat
     4741 ggtgaaaccc tgccgtctct actaaaaata caaaatttag ccaggtgtgg ctgtagtccc
     4801 agcgcggatc ccccagtgac gtaggaagtc catccattca cagcgcttct ataaaggcgc
     4861 cagctgaggc gcctactact ccaaccgcga ctgcagcgag caactccatg gtgagcaagg
     4921 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     4981 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     5041 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     5101 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     5161 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     5221 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     5281 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     5341 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     5401 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     5461 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     5521 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     5581 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     5641 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5701 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     5881 ttgtaagcgt taatcctact acacatataa aaggcgcgcc atcaacgtca aaacatgcac
     5941 acccaaaaaa
//
