LOCUS       sequence-313741-        6630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  AUTHORS   Shu T, Ayala R, Nguyen MD, Xie Z, Gleeson JG, Tsai LH
  TITLE     Ndel1 operates in a common pathway with LIS1 and cytoplasmic dynein 
            to regulate cortical neuronal positioning.
  JOURNAL   Neuron. 2004 Oct 14. 44(2):263-77.
  PUBMED    15473966
REFERENCE   2  (bases 1 to 6630)
  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: "Neuron. 
            2004 Oct 14. 44(2):263-77."
COMMENT     Sequence Label: pcDNA3 Lis1
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1949..1967)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1950..1967)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1987..2004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1993..2217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2263..2691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2350..2369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2560..2581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2700..2720)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2705..3034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2885..3020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2947..2966
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3101..3895
                     /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(3155..3174)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3765..3784
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4069..4190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4106..4125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4160..4179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4239..4255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4239..4255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4252..4274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4263..4279
                     /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(4287..4293,4294..4311,4312..4317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4332..4353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4470..4487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4641..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5400..6191,6192..6260))
                     /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     6023..6042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6261..6365)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6440..6459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac catggtgctg tcccagagac aacgagatga actaaatcga
      721 gctatagcag attatcttcg ttcaaatggc tatgaagagg catattccgt ttttaaaaag
      781 gaagctgaac tagatatgaa tgaagaatta gataagaagt atgctggtct tttggaaaaa
      841 aaatggacat ctgttattag attacaaaaa aaggtaatgg aattagaatc aaaactaaat
      901 gaagcaaaag aagaatttac gtcgggtggt cctcttggtc agaaacggga cccaaaagaa
      961 tggattcccc gtccacctga gaaatacgca ttgagtggtc ataggagtcc agttactcga
     1021 gtcattttcc atcctgtgtt cagtgttatg gtctctgctt cagaggatgc tacaattaag
     1081 gtgtgggatt atgagactgg agattttgag cgaactctca agggccatac agactctgta
     1141 caggacattt cctttgacca cagtggcaag cttctggctt cctgttcagc agatatgacg
     1201 attaaattat gggattttca gggttttgaa tgcatcagaa ccatgcacgg tcacgatcac
     1261 aatgtctctt cagtagccat catgcctaat ggagatcata tagtgtctgc ctcaagggat
     1321 aaaactataa agatgtggga agtgcaaact ggctactgtg tgaagacatt cacaggacac
     1381 agagaatggg tacgtatggt gcggccaaat caggatggca ctctgatagc cagctgttcc
     1441 aatgaccaga ctgtgcgtgt gtgggttgta gcaacaaagg aatgcaaggc tgagctccga
     1501 gaacatgaac atgtggtgga atgcatttcc tgggctccag aaagttcata ttcttccatc
     1561 tctgaagcaa caggatctga gactaaaaaa agtggcaagc ctggaccttt cttgctatct
     1621 ggttccagag acaaaactat taagatgtgg gacgtcagta ctggcatgtg ccttatgact
     1681 cttgtgggtc atgataactg ggtacgtgga gttctgttcc attctggggg gaagtttatt
     1741 ttgagttgtg ctgatgacaa gaccctccgt gtatgggatt ataagaacaa gcgatgcatg
     1801 aagaccctca atgcgcatga acactttgtt acctccttgg atttccataa gacggcaccc
     1861 tatgtggtta ctggcagtgt agatcaaaca gtaaaggtgt gggagtgccg ttgagcggcc
     1921 gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     1981 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2041 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2101 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2161 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2221 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2401 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2941 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3001 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3061 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3121 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3181 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3241 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3301 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3361 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3421 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3481 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3541 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3601 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3661 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3721 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3781 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3841 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3901 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3961 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4021 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4081 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4141 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4201 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4261 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4321 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4441 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6361 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6421 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6481 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6541 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     6601 ttcgcgatgt acgggccaga tatacgcgtt
//
