LOCUS       sequence-386300-        6414 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expresses TMSB10 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  TITLE     Tmsb10 triggers fetal Leydig differentiation by suppressing the 
            RAS/ERK pathway.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: p3xFLAG-TMSB10
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     130..152
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     144..161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     145..161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        322..701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        702..905
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     856..876
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..1000
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1177..1799
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1376..1395)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1823..1843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1841..2138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2192..2986
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2246..2265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2856..2875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3638..3772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3675..3694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3729..3748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3777..3796)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3778..3796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3810..3826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3810..3826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3823..3845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3834..3850
                     /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(3858..3864,3865..3882,3883..3888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3903..3924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4041..4058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4212..4800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4292..4311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4971..5762,5763..5831))
                     /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     5594..5613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5832..5936)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5963..6391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6050..6069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6260..6281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
       61 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      121 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat
      181 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
      241 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
      301 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      361 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      421 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      481 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      601 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      721 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      781 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      841 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      901 gagctcgttt agtgaaccgt cagaattaac catggactac aaagaccatg acggtgatta
      961 taaagatcat gacatcgatt acaaggatga cgatgacaag cttatggcag acaagccgga
     1021 catgggggaa atcgccagct tcgataaggc caagctgaag aaaaccgaga cgcaggagaa
     1081 gaacaccctg ccgaccaaag agaccattga acaggaaaag aggagtgaaa tctcctaaag
     1141 atctgatatc ggtaccagtc gactctagag gatcccgggt ggcatccctg tgacccctcc
     1201 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     1261 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     1321 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     1381 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     1441 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     1501 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     1561 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     1621 caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag
     1681 gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt
     1741 tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg
     1801 ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1861 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1981 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2041 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2101 tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc
     2161 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2221 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2281 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2341 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2401 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2461 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2521 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2581 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2641 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2701 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2761 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2821 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2881 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2941 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3001 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3061 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3121 cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt
     3181 tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc
     3241 gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag
     3301 tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag
     3361 aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg
     3421 cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa
     3481 gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat
     3541 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3601 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct
     3781 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     3841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4021 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5941 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6121 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6241 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6301 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6361 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct taca
//
