LOCUS       sequence-78278-e        5592 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 5592)
  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 5592)
  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-let-7g
FEATURES             Location/Qualifiers
     source          1..5592
                     /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(1144..1162)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1145..1162)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1182..1199)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1188..1412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1458..1886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1545..1564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1755..1776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1895..1915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1900..2230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2081..2216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2143..2162
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2297..3091
                     /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(2351..2370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2961..2980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3265..3386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3302..3321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3356..3375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3435..3451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3435..3451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3448..3470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3459..3475
                     /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(3483..3489,3490..3507,3508..3513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3528..3549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3666..3683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3837..4425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3917..3936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4596..5387,5388..5456))
                     /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     5219..5238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5457..5561)
                     /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 cccctgtctc aagtgcatcc tgaagagttc ctccagcgct ccgtttcctt
      961 ttgcctgatt ccaggctgag gtagtagttt gtacagtttg agggtctatg ataccacccg
     1021 gtacaggaga taactgtaca ggccactgcc ttgccaggaa cagcgcgcca gctgccaagt
     1081 ggggctgaga ggatggcgtc accctgctca tctctgctcg agcatgcatc tagagggccc
     1141 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     1201 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1261 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1321 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1381 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1441 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1801 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     1861 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     1921 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     1981 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2041 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2101 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2161 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2221 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     2281 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2341 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2401 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2461 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2521 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2581 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2641 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2701 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2761 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2821 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2881 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2941 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3001 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3061 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3121 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3181 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3241 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3301 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3361 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3421 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3481 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3541 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3601 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3661 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3721 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4021 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4261 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4501 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4561 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4621 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4681 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4741 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4801 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4861 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4921 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4981 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5041 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5101 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5161 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5221 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5281 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5341 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5401 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5461 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5521 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5581 ccacctgacg tc
//
