LOCUS       sequence-472652-        6777 bp    DNA     circular SYN 14-MAY-2025
DEFINITION  IVT mRNA of human gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6777)
  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 6777)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 14, 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: ARHGAP11B_pEF-DEST51
FEATURES             Location/Qualifiers
     source          1..6777
                     /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(3991..4015)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4046..4065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4047..4065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4082..5260)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4091..5029
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4114..4134)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     5548..5567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5642..5746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5747..5815,5816..6607)
                     /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(5965..5984)
                     /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 ttttttgttg aggtgtaata gaaggtgttc tgttaggttt
     3061 aaatttattt agtgctccac taacaaaatc tcctacacgt tgtcttctct ttctcttata
     3121 ttcaccagga gtttcatatt caccttcttc aaagccttcc agtgatggag tagcacagag
     3181 accatcaata cccaacatgg ctggtatctt ttccaggata aagtctggta cacgcccttc
     3241 ttttctgcgt tagaagacat cttttcatgt ccttcacttg tctgaagaag atttggtgca
     3301 aatattactg caagattgct gctatccatc ttattctcac tggatctaag agaaacattc
     3361 ctgagaaagt taaagaagta tcttaataca tgaactgtgt ggtcagccag aagacaggag
     3421 agcaacagta tagctttatt cttttcctct gtgcctaact gttgagcttt caaaagtgct
     3481 tcatgcaaat cagctgggag aatgggctct ggcagttccc taaaaaactg cttaagaagt
     3541 cccgcaatat cacaaggagg tgcagaagat aggcaacctt caccatgatc cactttattc
     3601 tttagtgctt ttaggcgaat cacagatcct gatttccgaa aaagcccttc ggtatgaata
     3661 tgttcttcta aagatgtgca agcatcgaca agaaagcttg gaatgtgtcc atattctggt
     3721 acagcagaat ggggcagtgc attaaaaggt actccaaata ttttaccccc tatttccgtg
     3781 gctgctgttt catgtctcct gcgatcgcac tgcccacgga cacccttcac cttaatacca
     3841 tagaaggccc gcagatgctg caacagggcc aacttcacca gcctctgatc ccacattccg
     3901 gatacgtcga taactctgag gcaggatgca ggtcctgacc ctcgttcgcc accaagtctt
     3961 ccaatttcca aacgctaagg gcgaattcgg agcctgcttt tttgtacaaa cttgttgatc
     4021 aagcttacct agccagcttg ggtctcccta tagtgagtcg tattagtacc aagctaattc
     4081 ctcacgacac ctgaaatgga agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga
     4141 accaagatcc aaactcaaaa agggcaaatt ccaaggagaa ttacatcaag tgccaagctg
     4201 gcctaacttc agtctccacc cactcagtgt ggggaaactc catcgcataa aacccctccc
     4261 cccaacctaa agacgacgta ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg
     4321 cccggtactc cgtggagtca catgaagcga cggctgagga cggaaaggcc cttttccttt
     4381 gtgtgggtga ctcacccgcc cgctctcccg agcgccgcgt cctccatttt gagctccctg
     4441 cagcagggcc gggaagcggc catctttccg ctcacgcaac tggtgccgac cgggccagcc
     4501 ttgccgccca gggcggggcg atacacggcg gcgcgaggcc aggcaccaga gcaggccggc
     4561 cagcttgaga ctacccccgt ccgattctcg gtggccgcgc tcgcaggccc cgcctcgccg
     4621 aacatgtgcg ctgggacgca cgggccccgt cgccgcccgc ggccccaaaa accgaaatac
     4681 cagtgtgcag atcttggccc gcatttacaa gactatcttg ccagaaaaaa agcgtcgcag
     4741 caggtcatca aaaattttaa atggctagag acttatcgaa agcagcgaga caggcgcgaa
     4801 ggtgccacca gattcgcacg cggcggcccc agcgcccagg ccaggcctca actcaagcac
     4861 gaggcgaagg ggctccttaa gcgcaaggcc tcgaactctc ccacccactt ccaacccgaa
     4921 gctcgggatc aagaatcacg tactgcagcc aggtggaagt aattcaaggc acgcaagggc
     4981 cataacccgt aaagaggcca ggcccgcggg aaccacacac ggcacttacc tgtgttctgg
     5041 cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc
     5101 ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc
     5161 gccaccttct ctaggcaccg gttcaattgc cgacccctcc ccccaacttc tcggggactg
     5221 tgggcgatgt gcgctctgcc cactgacggg caccggagcc aattcccact cctttcaaga
     5281 cctagaaggt ccattagctg caaagattcc tctctgttta aaactttatc catctttgca
     5341 aagctttttg caaaagccta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     5401 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     5461 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     5521 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5581 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     5641 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5701 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5761 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5821 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5881 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5941 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6001 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6061 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6121 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6181 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6241 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6301 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6361 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6421 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6481 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6541 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6601 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6661 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6721 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
