LOCUS       pUC57-5'H-2A-dTo        6953 bp DNA     circular SYN 28-OCT-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  AUTHORS   Adamson
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(146..396)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="LAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQL
                     RSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     primer_bind     357..413
                     /label=Fragment 1.FOR
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    397..1196
                     /label=5'H
     CDS             397..1196
                     /codon_start=1
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /product="transcription factor SOX-17"
                     /label=SOX17
                     /note="Derived by automated computational analysis using
                     gene prediction method: BestRefSeq."
                     /db_xref="CCDS:CCDS6159.1"
                     /db_xref="Ensembl:ENSP00000297316.4"
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
                     /protein_id="NP_071899.1"
                     /translation="KRVEGGFLHGLAEPQAAALGPEGGRVAMDGLGLQFPEQGFPAGPP
                     LLPPHMGGHYRDCQSLGAPPLDGYPLPTPDTSPLDGVDPDPAFFAAPMPGDCPAAGTYS
                     YAQVSDYAGPPEPPAGPMHPRLGPEPAGPSIPGLLAPPSALHVYYGAMGSPGAGGGRGF
                     QMQPQHQHQHQHQHHPPGPGQPSPPPEALPCRDGTDPSQPAELLGEVDRTEFEQYLHFV
                     CKPEMGLPYQGHDSGVNLPDSHGAISSVVSDASSAVYYCNYPDV"
     mRNA            397..1196
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /product="SRY-box 17"
                     /label=SOX17 mRNA
                     /note="Derived by automated computational analysis using
                     gene prediction method: BestRefSeq.
                     /transcript_id=NM_022454.4"
                     /db_xref="Ensembl:ENST00000297316.5"
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
     gene            397..1196
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /label=SOX17
                     /note="SRY-box 17; Derived by automated computational
                     analysis using gene prediction method: BestRefSeq."
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
     primer_bind     826..844
                     /label=NHa03 TIDE F
     primer_bind     complement(1165..1216)
                     /label=Fragment 1.REV
     primer_bind     1177..1212
                     /label=Fragment 2.FOR
     misc_feature    1181..1196
                     /label=NHa03 g078
                     /note="https://www.sanger.ac.uk/htgt/wge/crispr/1041108078"
     misc_feature    complement(1191..1196)
                     /label=NHa03 g079
                     /note="https://www.sanger.ac.uk/htgt/wge/crispr/1041108079"
     CDS             1197..1250
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A - rare codons changed
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1249..1254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1251..1952
                     /label=dTOMATO_reference-genbank_HQ418395_reference-g
                     /label=tdTOMATO_reference-genbank_HQ418395_reference-genba 
                     nk_ADC42843
     primer_bind     complement(1282..1301)
                     /label=NHa03 tdTomato R
     primer_bind     complement(1932..1975)
                     /label=Fragment 2.REV
     primer_bind     1933..1983
                     /label=Fragment 3.FOR
     protein_bind    complement(1989..2022)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            1989..2022
                     /label=loxP
                     /note="loxP"
     gene            2081..2595
                     /label=PGK promoter
                     /note="PGK promoter"
     promoter        2087..2586
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     gene            2598..2663
                     /label=EM7 promoter
                     /note="EM7 promoter"
     promoter        2598..2645
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2664..3467
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2664..3464
                     /label=Neo
                     /note="Neo"
     gene            3481..3783
                     /label=bGHpA
                     /note="bGHpA"
     polyA_signal    3505..3729
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3684..3703
                     /label=NHa03 bGH pA F
     protein_bind    complement(3786..3819)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            3786..3819
                     /label=loxP
                     /note="loxP"
     primer_bind     complement(3826..3871)
                     /label=Fragment 3.REV
     primer_bind     3832..3867
                     /label=Fragment 4.FOR
     primer_bind     complement(3835..3851)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mRNA            3850..4651
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /product="SRY-box 17"
                     /label=SOX17 mRNA
                     /note="Derived by automated computational analysis using
                     gene prediction method: BestRefSeq.
                     /transcript_id=NM_022454.4"
                     /db_xref="Ensembl:ENST00000297316.5"
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
     gene            3850..4651
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /label=SOX17
                     /note="SRY-box 17; Derived by automated computational
                     analysis using gene prediction method: BestRefSeq."
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
     misc_feature    complement(3850..3862)
                     /label=NHa03 g079
                     /note="https://www.sanger.ac.uk/htgt/wge/crispr/1041108079"
     misc_feature    3850..3852
                     /label=NHa03 g078
                     /note="https://www.sanger.ac.uk/htgt/wge/crispr/1041108078"
     CDS             3850..3851
                     /codon_start=1
                     /gene="SOX17"
                     /gene_synonym="VUR3"
                     /product="transcription factor SOX-17"
                     /label=SOX17
                     /note="Derived by automated computational analysis using
                     gene prediction method: BestRefSeq."
                     /db_xref="CCDS:CCDS6159.1"
                     /db_xref="Ensembl:ENSP00000297316.4"
                     /db_xref="GeneID:64321"
                     /db_xref="HGNC:HGNC:18122"
                     /db_xref="MIM:610928"
                     /protein_id="NP_071899.1"
                     /translation=""
     misc_feature    3852..4651
                     /label=3'H
     primer_bind     complement(4174..4193)
                     /label=NHa03 TIDE R
     primer_bind     complement(4622..4691)
                     /label=Fragment 4.REV
     CDS             complement(4652..4736)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPISRCIREV"
     primer_bind     complement(4732..4748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4756..4772
                     /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(4780..4786,4787..4804,4805..4810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4825..4846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5134..5722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5893..6684,6685..6753))
                     /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"
     promoter        complement(6754..6858)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgtgaa gcgggtggag ggcggcttcc
      421 tgcacggcct ggctgagccg caggcggccg cgctgggccc cgagggcggc cgcgtggcca
      481 tggacggcct gggcctccag ttccccgagc agggcttccc cgccggcccg ccgctgctgc
      541 ctccgcacat gggcggccac taccgcgact gccagagtct gggcgcgcct ccgctcgacg
      601 gctacccgtt gcccacgccc gacacgtccc cgctggacgg cgtggacccc gacccggctt
      661 tcttcgccgc cccgatgccc ggggactgcc cggcggccgg cacctacagc tacgcgcagg
      721 tctcggacta cgctggcccc ccggagcctc ccgccggtcc catgcacccc cgactcggcc
      781 cagagcccgc gggtccctcg attccgggcc tcctggcgcc acccagcgcc cttcacgtgt
      841 actacggcgc gatgggctcg cccggggcgg gcggcgggcg cggcttccag atgcagccgc
      901 aacaccagca ccagcaccag caccagcacc accccccggg ccccggacag ccgtcgcccc
      961 ctccggaggc actgccctgc cgggacggca cggaccccag tcagcccgcc gagctcctcg
     1021 gggaggtgga ccgcacggaa tttgaacagt atctgcactt cgtgtgcaag cctgagatgg
     1081 gcctccccta ccaggggcat gactccggtg tgaatctccc cgacagccac ggggccattt
     1141 cctcggtggt gtccgacgcc agctccgcgg tatattactg caactatcct gacgtggagg
     1201 gcaggggaag cctgctgaca tgcggggacg tggaggaaaa tcccggcccc atggtgagca
     1261 agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga
     1321 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1381 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1441 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca
     1501 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1561 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga
     1621 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     1681 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc
     1741 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg
     1801 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct
     1861 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc
     1921 tgttcctgta cggcatggac gagctgtaca agtgagccca attccgatca tattcaataa
     1981 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtctg aagaggagtt
     2041 tacgtccagc caagctagct tggctgcagg tcgtcgaaat tctaccgggt aggggaggcg
     2101 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
     2161 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
     2221 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
     2281 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
     2341 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
     2401 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
     2461 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
     2521 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
     2581 ctttcgacct gcagcctgtt gacaattaat catcggcata gtatatcggc atagtataat
     2641 acgacaaggt gaggaactaa accatgggat cggccattga acaagatgga ttgcacgcag
     2701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2821 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3181 tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg acccatggcg
     3241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg gatcaattct
     3481 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3541 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3601 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3661 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3721 gctctatggc ttctgaggcg gaaagaacca gctggggctc gactagagct tgcggaaccc
     3781 ttaatataac ttcgtataat gtatgctata cgaagttatt aggtccctcg aggggatcca
     3841 ctagttctag acaggtccct gatccgcccc agcctgcagg ccagaagcag tgttacacac
     3901 ttcctggagg agctaaggaa atcctcagac tcctgggttt ttgttgttgc tgttgttgtt
     3961 ttttaaaagg tgtgttggca tataatttat ggtaatttat tttgtctgcc acttgaacag
     4021 tttggggggg tgaggtttca tttaaaattt gttcagagat ttgtttccca tagttggatt
     4081 gtcaaaaccc tatttccaag ttcaagttaa ctagctttga atgtgtccca aaacagcttc
     4141 ctccatttcc tgaaagttta ttgatcaaag aaatgttgtc ctgggtgtgt tttttcaatc
     4201 ttctaaaaaa taaaatctgg aatcctgctt ttttgctcta ctagtacctc tgtcacacta
     4261 gtcttatcaa aaaccagttc ttaagatcaa tgttaagttt attagttaat gtaaatttct
     4321 catcctcgaa aagggtgaac ataaatgcct ttaaggagta tatctaaaaa taaacattag
     4381 gatatctaag tttgatgtaa ttgtttcagg aaggaaaaaa gaaaagcatt ctggaatgag
     4441 cctacttcaa gtaatcttag tttctaaaac taacagttaa tattttcaat tccagtatat
     4501 cactttaagt agaaggggat gtccaagtaa ttttggtttt ctaactgttg aatcataagc
     4561 ttgacctgcc cccagaggct ttttggatgt ttttatctgt gttttgccat ctctttacac
     4621 tcctcgacat tcagtttacc ttaatcttca cgtacctcgc gaatgcatct agatatcgga
     4681 tcccgggccc gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct
     4741 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4801 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5641 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5701 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5761 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5821 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6841 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6901 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
