LOCUS       sequence-474310-        5525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT mRNA of human gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5525)
  AUTHORS   Soto DC, Uribe-Salazar JM, Kaya G, Valdarrago R, Sekar A, Haghani 
            NK, Hino K, La GN, Mariano NAF, Ingamells C, Baraban AE, Turner TN, 
            Green ED, Simo S, Quon G, Andres AM, Dennis MY
  TITLE     Gene expansions contributing to human brain evolution.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.09.26.615256. doi: 
            10.1101/2024.09.26.615256.
  PUBMED    39386494
REFERENCE   2  (bases 1 to 5525)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.09.26.615256. doi: 
            10.1101/2024.09.26.615256."
COMMENT     Sequence Label: PTPN20CP_pGCS1
FEATURES             Location/Qualifiers
     source          1..5525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1143..1277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1180..1199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1234..1253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1307..1331
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2737..2761)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2801..2819)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2802..2819)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2854..3838)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3909..4364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3996..4015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4206..4227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4390..4494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4495..4563,4564..5355)
                     /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(4713..4732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaat
     1081 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1141 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1261 cttatcatgt ctggatctac gtaatacgac tcactatagt tctagaacca ctttgtacaa
     1321 gaaagctggg tctgtgaaag gatcgagaca tccacacagc agactgaact gcaaaagcat
     1381 tggcatttct cttctgggca ttttcttcgt ctcctagctg gagatcctgc agatgccact
     1441 tctttttctt tgcagctttg ggtccaccgt ttttctttct gtgtttcttc tggtccttag
     1501 tctccatagt ggctatttac cgcagtgaac agttctttac gatggcacag aacacgacat
     1561 ccacacatag gaacacccct gtccggccta tgccggcact gcagtgaaca accatgggtc
     1621 ctgtaaggtg gctcttcctt gcataacgaa tatattttat gaagctatct gctgaggcag
     1681 gagtgccatg gtctggccac ttggtgaact gcaactgttt tacagagtga ctagttcccg
     1741 tggacttctc cacaacttga aacattcgaa tgatgaaata ttgaagtatc tggtagttct
     1801 ccaggaatac acggaagtgt ttcaattcca atggcttctt cagagaaatg ggccagtaat
     1861 ggtagcattt gataattcca ccttctatct ctctggttat catggcaata acattcgaat
     1921 tattttccaa caccatttgc caaaagtcat ctatggtgct cagcagtggt ccttgagtag
     1981 cgatataaaa atactcttct ccacaattga ctattctaat ataactagca ttgatgtagt
     2041 ccttgctttt tccaagagga acgcgtgttg aatcatatgg aagaatatct cggtatctgt
     2101 ttttttctct gttgcttggt tgattcccag agttgaactc accaggcaga ttcttaagtt
     2161 ctaaagccat aaattcctgc atgatatcat acgctgctgt tttttcctca aggattttca
     2221 aacaatcctt gatggctgtc tgctcatgaa aattgaatat taatggccga atctgagcca
     2281 actgatctag ttctccttca gaaacaattt tgcgcgtatc agaaagtaga ggggagagag
     2341 cctgtgatgg ccctgcagcc tcctcatcct cactgctcca cctgtctctt ccgaaggggc
     2401 ctctggctgt ccacatgctg ggatcactgc cactttctga aggctcttca aaaacatcct
     2461 cataatcatt atgtgggaaa ttccgaagag aaagttttac cttctctgaa ttaactttat
     2521 tttcaaaaac ctttgatcta ggtgtgtttt cctgacttga tgaaggcaaa gcctccctga
     2581 aattcaggtc ttctgcttca gagtcatttc cctcataatc gtttacagac tctgctctaa
     2641 agtccctagg tgaagacatg ttgttcccat cccctaaatg agactggtaa aaggcttctt
     2701 cagaaaagga gccttagtgt cctgtgagtc aggccaagcc tgcttttttg tacaaacttg
     2761 tggccggccg gatcctgcaa aaagaacaag tagcttgtat tctatagtgt cacctaaatc
     2821 aagcttgctc cgacgtcccc aggcagaatg gcggttccct aaacgagcat tgcttatata
     2881 gacctcccat taggcacgcc taccgcccat ttacgtcaat ggaacgccca tttgcgtcat
     2941 tgcccctccc cattgacgtc aatggggatg tacttggcag ccatcgcggg ccatttaccg
     3001 ccattgacgt caatgggagt actgccaatg taccctggcg tacttccaat agtaatgtac
     3061 ttgccaagtt actattaata gatattgatg tactgccaag tgggccattt accgtcattg
     3121 acgtcaatag ggggcgtgag aacggatatg aatgggcaat gagccatccc attgacgtca
     3181 atggtgggtg gtcctattga cgtcaatggg cattgagcca ggcgggccat ttaccgtaat
     3241 tgacgtcaat gggggaggcg ccatatacgt caataggacc gcccatatga cgtcaatagg
     3301 aaagaccata tatagagacc attgacgtca atgggggagt ggctatgggc ggtattagga
     3361 agccccatat atggtatatg ggaccgccca ttgggagggg ctatctacgt caataggaaa
     3421 acccatatat ggaatactat atggcatagg gccaatacat agtattgaac ctggccaata
     3481 gccatattgg catagggcca tattggatat tgcctatata ttgatcctgg catatagcca
     3541 atatggccgc cattattggc accatgccaa ttcaatatgg cggacttggc acagggccaa
     3601 gacccctcct caaacttggc accgtgccaa gtagacccct ccccagttgg cacagtgcca
     3661 ggtccgccat attgaattgg catggtgcca agtccgccat attgaattgg ctggggtcca
     3721 gatccaccat attgaattgg cacgagtcca tatacgccat attgaattgg cacaggtcca
     3781 gatccaccat attgaattgg catgagtcca tatacgccat attgaattgg cggtcgactg
     3841 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3901 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4381 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttc
//
