LOCUS       sequence-326375-        8321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry2-expression vector for human H11 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  AUTHORS   Pryzhkova MV, Jordan PW
  TITLE     Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell
            and Testis Development Research.
  JOURNAL   Tissue Eng Regen Med. 2020 Apr;17(2):223-235. doi: 
            10.1007/s13770-020-00240-0. Epub 2020 Feb 29.
  PUBMED    32114677
REFERENCE   2  (bases 1 to 8321)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s13770-020-00240-0"; journalName: "Tissue Eng Regen Med"; 
            date: "2020-04"; volume: "17"; issue: "2"; pages: "223-235"
COMMENT     Sequence Label: p273 EF1a_mCherry2_hH11
FEATURES             Location/Qualifiers
     source          1..8321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..27
                     /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(35..41,42..59,60..65))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    80..101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(218..235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(389..977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(469..488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1148..1939,1940..2008))
                     /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     1771..1790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2009..2113)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2140..2595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2227..2246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2437..2458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2721..2743
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2735..2752
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2736..2752
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3568..4746
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3799..4737
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4694..4714
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4760..4769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4766..5476
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4911..4929)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    5594..5715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5631..5650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5685..5704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5756..5789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5826..6325
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5861..5884)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6258..6277
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6365..6964
                     /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(6365..6384)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6861..6881
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(7545..7578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
       61 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      181 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     2161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2341 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2401 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2461 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2521 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2581 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     2641 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     2701 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     2761 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     2821 aatcaatatt ggctattggc cattgcatac gtttcttgtg aactcatctc gtgcctgcaa
     2881 gacatcaaaa gtttctccaa agacatccct tcctctctgc cctcatccta tatcaaggtc
     2941 ttagctcctt gaagactgca ttaatgtgtc ttttgccttc atcccccccc caccctcccg
     3001 cttcctctag tccttgtgtt taccaaaata cttttgctaa aacctgtatg tctagcttct
     3061 gctgatactc ttagcaatac tcccttattt ccttctggcg ccatttgcca atcaccagct
     3121 aatggctttg ctctttgcat ggtacctgtt tctgtcctac tagcttctag tctgaaaagg
     3181 aagcctttac tagcagcttc aacccagtcc tccttacctt gcacctatta acgttggttc
     3241 aacatgaagg ggaagtaaga taagtggaag aggatacaca ggaaaggtgg aactggattc
     3301 aggtatctgt gttctgccat tgacacttgg gactgcgttc cagcttccac ctgtggaaac
     3361 cagaaagcat ttgccaaaaa ttctaaacag agggtctatt tctgaagctg aggaatcaca
     3421 tggagtgaat agcatggggg aaggggatta ctaatgaggc tgaaagaaaa gtaccaggaa
     3481 gagactgact tcatgcataa tccataaagc caggttttct tgaaaggtga cctgctttct
     3541 tgtattcctg caagaaacgg ctcgaggggc tccggtgccc gtcagtgggc agagcgcaca
     3601 tcgcccacag tccccgagaa gttgtgggga ggggtcggca attgaagcgg tgcctagaga
     3661 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3721 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3781 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
     3841 ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat
     3901 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
     3961 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg
     4021 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
     4081 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
     4141 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
     4201 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
     4261 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     4321 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     4381 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     4441 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     4501 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     4561 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     4621 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     4681 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     4741 tcgtgagggg atcgtcgacg ccaccatggt gagcaagggc gaggaggata acatggccat
     4801 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     4861 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     4921 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     4981 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     5041 ccccgagggc ttcaattggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     5101 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     5161 caacttcccc tccgacggcc ccgtaatgca gtgtcgtacc atgggctggg aggcctccac
     5221 tgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     5281 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     5341 gcagctgccc ggcgcctaca acgtcgacat caagttggac atcctttccc acaacgagga
     5401 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     5461 cgagctgtac aagtaatcta gataactgat cataatcagc cataccacat ttgtagaggt
     5521 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5581 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5701 catcaatgta tcttaacgcg atcgccagta ctagtgaacc tcttcgaggg acctaataac
     5761 ttcgtatagc atacattata cgaagttata ttaagggttc cggatctcga cctcgaaatt
     5821 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     5881 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     5941 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     6001 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     6061 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     6121 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     6181 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     6241 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     6301 ctcttcctca tctccgggcc tttcgacctg catccatcta gatctcgagc agctgaagct
     6361 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     6421 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     6481 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     6541 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     6601 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     6661 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     6721 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     6781 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     6841 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     6901 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     6961 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     7021 gatgatatca gatccccggg atgcagaaat tgatgatcta ttaaacaata aagatgtcca
     7081 ctaaaatgga agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca
     7141 ttttgaatgg aaggattgga gctacggggg tgggggtggg gtgggattag ataaatgcct
     7201 gctctttact gaaggctctt tactattgct ttatgataat gtttcatagt tggatatcat
     7261 aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca tgatctatag
     7321 atctatagat ctctcgtggg atcattgttt ttctcttgat tcccactttg tggttctaag
     7381 tactgtggtt tccaaatgtg tcagtttcat agcctgaaga acgagatcag cagcctctgt
     7441 tccacataca cttcattctc agtattgttt tgccaagttc taattccatc agaagctggt
     7501 cgagatccta agcttggctg gacgtaaact cctcttcaga cctaataact tcgtatagca
     7561 tacattatac gaagttatat taagggttat tgaatatgat cggaattgcg gccctgacct
     7621 gttggggtct ttaaagctca aggaaaaagg ccatagttga tttctcctaa atcaagatag
     7681 agtccaatta actttttttt ttttttaaga tggagtctca ctctgtcgcc aggctggact
     7741 gcagtggcgt gatctcagct cactgcaacc tctgcctccc aggttcaggc gattcttctg
     7801 tctcagcctc ccgagtagct gggactacaa gtgtgggcca ccatgcccag ctagttttgt
     7861 atttttagta gagatgggat ttcaccatct tggctaggat ggtcttgatc tcttgacctt
     7921 gtgattcgcc tgcctctgcc tcccagagtg ctgagattac aggcgtgagc cacagtgccc
     7981 aatttccatg ggttttcaag aaaaacttaa cttacttaaa cttcagatca cttccttaaa
     8041 ggaacatact gagcatagct tggctggtta gaaattagga agactagctt gggctacatg
     8101 gtaagaccct gcctctacaa aaaataagaa aaaagttagt ggcatgtgcc tgtagttcca
     8161 tctacttggg aggctgaggt gagaggatcg cttgagccca ggaggttgaa gctgcagtga
     8221 gccacgattg caccactgta ctccagcctg ggtgacacag agtgagaccc tgtctccaaa
     8281 caaaattagg aagggtttca gagaggaaat aaacacggcc g
//
