LOCUS       sequence-472673-        7242 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 7242)
  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 7242)
  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: SRGAP2C_pEF-DEST51
FEATURES             Location/Qualifiers
     source          1..7242
                     /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     732..749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    866..887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(902..907,908..925,926..932)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    940..956
                     /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     945..967
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     964..980
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     964..980
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1029..1150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1040..1059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1094..1113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1308..1706)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(1725..1772)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1820..2149)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1834..1969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1888..1907)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2134..2154
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2163..2591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2273..2294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2485..2504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2637..2861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2850..2867
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2890..2907)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2917..2958)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    2984..3008
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4456..4480)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4511..4530)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4512..4530)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4547..5725)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4556..5494
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4579..4599)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     6013..6032
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6107..6211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6212..6280,6281..7072)
                     /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(6430..6449)
                     /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 ctcacatgtt ctttcctgcg ttatcccctg
      661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      781 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      841 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      901 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      961 acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     1021 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1081 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1141 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     1201 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     1261 cgaaatctcg tagcacgtgt cagtcctgct cctcggccac gaagtgctta gccctcccac
     1321 acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc
     1381 ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag
     1441 gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct
     1501 gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga
     1561 tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg
     1621 tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat
     1681 tcttcttgag acaaaggctt ggccatggtt tagttcctca ccttgtcgta ttatactatg
     1741 ccgatatact atgccgatga ttaattgtca acacgtgctg atcagatccg aaaatggata
     1801 tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     1861 tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     1921 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     1981 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     2041 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     2101 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa
     2161 ttaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2221 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2281 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2341 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2461 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2521 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2581 acagggcgcg tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat
     2641 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2701 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2761 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2821 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt
     2881 ttaaactcaa tggtgatggt gatgatgacc ggtacgcgta gaatcgagac cgaggagagg
     2941 gttagggata ggcttacctt cgaaccgcgg gccctctaga tcaaccactt tgtacaagaa
     3001 agctgggtcg aattcgccct tcagtgcatg ctgacgctct ttaaaagcca tagcactccc
     3061 ttactgtgaa ataaaactgc tctgtctctt gctggttggc tctcctctta gcaatgctgg
     3121 gcttgctcat gaaggtttca gagaccgtgg acttgacgga ctccatggag ttgctgtact
     3181 ggaagcagtc agacacatca aagtcctcga cagtcacaat gtcctggatg gtttgcaggg
     3241 tggcctccat tgtcttcttt acctcttcgt tttcaatctt tagagtggat aagcgagact
     3301 gcagttgttg gcatctctgt accagctcac tctggacagg ctgctgggca cagagctggg
     3361 aagccatatc ccccatgtgg ggctgaaact caaacttcat aggggggcag aagacgttgt
     3421 tgtacatctc catgaggcgc tgcttgtcac tggtggcatc caggttttct actgcattct
     3481 cgatggcatc cagaccctca tgcttcgact gttccaggtt taactcagca gagaggaagg
     3541 tgcgtagagc ccggttcaga cttgcatggt agcctaagtc acaacaatca ataaggtcag
     3601 ataggtcatg gatgtagtac ttgaagacag atgcattggt tgcctccaaa gccagcaagt
     3661 actcattctg ggctttgatg gccttcagct tattctccgt gtacttggct tggcgcttct
     3721 ccttcatctt ctcaatcttc ttcactgagc tcctccggac atgtttctcc tcaatgcgaa
     3781 cattggccgt ggagtcaggg gagcgtgggg tctgccggtc ctcctgcttt accgatttac
     3841 caatttgctt ctcctcctgc ttctccgcct cctttagttt gctctgagca ctgatgctgt
     3901 cggcattgta catgtgatat gtcttcatga ccgagtagag ctcgttcagg accttcatca
     3961 aatcatcttg gagctgctgg ccgacttctt tactcttttt aaagagtctt cctgagtcct
     4021 cgctgacttg tacaaatcga ggaatgatat tattcaggta gatgtcactc agggtggtat
     4081 ggtccctgct ttcccacttc acctggttta agaggagatt ccagcagttg actggagaga
     4141 gaacattctg atccttcttg aattgctggt ccttggtgct gcgtgtcttg gccaggaagt
     4201 gttctgccag cttctccagg ttgcgggagt agtccatctc aatctctgcc ttctttcgga
     4261 agaagtcctg gaggtcctgc aacagttgca cccgaagctc acactgctgg tccaggcatt
     4321 tcatctgctc tgtgagctga gcacggatct ctttgacctg agtatcgtac tctgctatga
     4381 tctccttatc ctttttgaat ttggctggag acgtcatgtt ttccttcttt cttccaaatt
     4441 caagggcgaa ttcggagcct gcttttttgt acaaacttgt tgatcaagct tacctagcca
     4501 gcttgggtct ccctatagtg agtcgtatta gtaccaagct aattcctcac gacacctgaa
     4561 atggaagaaa aaaactttga accactgtct gaggcttgag aatgaaccaa gatccaaact
     4621 caaaaagggc aaattccaag gagaattaca tcaagtgcca agctggccta acttcagtct
     4681 ccacccactc agtgtgggga aactccatcg cataaaaccc ctccccccaa cctaaagacg
     4741 acgtactcca aaagctcgag aactaatcga ggtgcctgga cggcgcccgg tactccgtgg
     4801 agtcacatga agcgacggct gaggacggaa aggccctttt cctttgtgtg ggtgactcac
     4861 ccgcccgctc tcccgagcgc cgcgtcctcc attttgagct ccctgcagca gggccgggaa
     4921 gcggccatct ttccgctcac gcaactggtg ccgaccgggc cagccttgcc gcccagggcg
     4981 gggcgataca cggcggcgcg aggccaggca ccagagcagg ccggccagct tgagactacc
     5041 cccgtccgat tctcggtggc cgcgctcgca ggccccgcct cgccgaacat gtgcgctggg
     5101 acgcacgggc cccgtcgccg cccgcggccc caaaaaccga aataccagtg tgcagatctt
     5161 ggcccgcatt tacaagacta tcttgccaga aaaaaagcgt cgcagcaggt catcaaaaat
     5221 tttaaatggc tagagactta tcgaaagcag cgagacaggc gcgaaggtgc caccagattc
     5281 gcacgcggcg gccccagcgc ccaggccagg cctcaactca agcacgaggc gaaggggctc
     5341 cttaagcgca aggcctcgaa ctctcccacc cacttccaac ccgaagctcg ggatcaagaa
     5401 tcacgtactg cagccaggtg gaagtaattc aaggcacgca agggccataa cccgtaaaga
     5461 ggccaggccc gcgggaacca cacacggcac ttacctgtgt tctggcggca aacccgttgc
     5521 gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa
     5581 aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg
     5641 caccggttca attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct
     5701 ctgcccactg acgggcaccg gagccaattc ccactccttt caagacctag aaggtccatt
     5761 agctgcaaag attcctctct gtttaaaact ttatccatct ttgcaaagct ttttgcaaaa
     5821 gcctagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa
     5881 aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag
     5941 cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact
     6001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag
     6061 atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6181 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6241 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6301 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6361 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6421 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6481 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6541 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6601 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6661 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6721 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6781 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6841 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6901 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6961 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7021 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7081 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7141 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7201 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
