LOCUS       sequence-78279-e        6466 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 6466)
  AUTHORS   Heo I, Joo C, Cho J, Ha M, Han J, Kim VN
  TITLE     Lin28 mediates the terminal uridylation of let-7 precursor MicroRNA.
  JOURNAL   Mol Cell. 2008 Oct 24;32(2):276-84. doi: 
            10.1016/j.molcel.2008.09.014.
  PUBMED    18951094
REFERENCE   2  (bases 1 to 6466)
  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"; doi: 
            "10.1016/j.molcel.2008.09"; journalName: "Mol Cell"; date: 
            "2008-10-24- 24"; volume: "32"; issue: "2"; pages: "276-84"
COMMENT     Sequence Label: pcDNA3-pri-miR-16-1
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2018..2036)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2019..2036)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2056..2073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2062..2286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2332..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2629..2650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2769..2789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2774..3104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2955..3090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3017..3036
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3171..3965
                     /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(3225..3244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3835..3854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4139..4260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4230..4249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4309..4325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4309..4325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4322..4344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4333..4349
                     /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(4357..4363,4364..4381,4382..4387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4402..4423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4540..4557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4711..5299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4791..4810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5470..6261,6262..6330))
                     /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     6093..6112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6331..6435)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgattcat ggatcccctt
      961 ggagtaaagt agcagcacat aatggtttgt ggattttgaa aaggtgcagg ccatattgtg
     1021 ctgcctcaaa aatacaagga tctgatcttc tgaagaaaat atatttcttt ttattcatag
     1081 ctcttatgat agcaatgtca gcagtgcctt agcagcacgt aaatattggc gttaagattc
     1141 taaaattatc tccagtatta actgtgctgc tgaagtaagg ttgaccatac tctacagttg
     1201 tgttttaatg tatattaatg ttactaatgt gttttcagtt ttattgatag tcttttcagt
     1261 attattgata atcttgttat ttttagtatg attctgtaaa aatgaattaa tactaatttt
     1321 tcagatgtat catctcttaa aatactgtaa ttgcaattta ataattgtat tgaatgccat
     1381 caagtttttt taaaaagctt atgcagcatt agaggaattt attttaatgc acatttatat
     1441 tcaacataga cattaattca gatttttact tgggataaaa caaattctag ttttcccttt
     1501 gttttgaaat tacttttaaa atatgtcttt acagataaat ataaaatata ttaagcattt
     1561 tgaacagagc ttagaagaca atatttagta ctgtttctga atatttcttt atatctgaag
     1621 gggaaaagcc atcaaaatat gtgaattaaa tacctaaaat tctggttgtc aaaacgtcac
     1681 acttaaccat aactttaaag gtagaaaaac ccttacaagt gaccaaccca ctcattggat
     1741 agctaaggaa acacaaaact caaataattt gctacacatc acagattggg gtcttagtct
     1801 tatgtcttct aactttataa tggtatacta tttttttcat gtttttatta aaatgttttc
     1861 ttaaatcctt agccaaagat ttttgtttga taagtcaaaa ataaggtctt gtttactttg
     1921 tttacttttt tgtattcaag atttaactca gagtaaggaa ttctgcagat atccatcaca
     1981 ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta
     2041 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2101 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2161 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2221 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2281 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2341 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2401 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2461 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     2521 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2581 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2641 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2701 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2761 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca
     2821 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2881 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2941 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3001 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3061 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3121 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3181 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3241 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3301 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3361 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3421 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3481 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3541 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3601 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3661 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3721 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3781 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3841 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3901 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3961 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4021 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4081 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4261 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4321 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4381 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4441 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4501 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4561 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4621 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     4921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6361 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6421 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
