LOCUS       40924_37488        6380 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6380)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        192..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             744..1460
                     /codon_start=1
                     /label=AcGFP1
                     /note="Aequorea coerulescens GFP"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILL
                     EFVTAAGITHGMDELYK"
     polyA_signal    1478..1599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1700..2128
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2142..2471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2538..3329
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3506..3639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3676..3692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3700..3716)
                     /label=lac operator
                     /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(3724..3754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3769..3790)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4078..4663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4837..5694)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5695..5799)
                     /label=AmpR promoter
     promoter        6065..6083
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6214..6380
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
        1 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg gtttatatat
       61 cttgtggaaa ggacgcggga tccgagctcg gtaccaagct taagtttaaa ccgctgatca
      121 gcctcgactg tgccttctaa atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      721 cgctagcgct accggtcgcc accatggcca gcaagggcga ggagctgttc accggcgtgg
      781 tgcccatcct ggtggagctg gacggcgatg tgaatggcca caagttcagc gtgagcggcg
      841 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      901 agctgcctgt gccctggccc accctggtga ccaccctgag ctacggcgtg cagtgcttct
      961 cacgctaccc cgatcacatg aagcagcacg acttcttcaa gagcgccatg cctgagggct
     1021 acatccagga gcgcaccatc ttcttcgagg atgacggcaa ctacaagtcg cgcgccgagg
     1081 tgaagttcga gggcgatacc ctggtgaatc gcatcgagct gaccggcacc gatttcaagg
     1141 aggatggcaa catcctgggc aataagatgg agtacaacta caacgcccac aatgtgtaca
     1201 tcatgaccga caaggccaag aatggcatca aggtgaactt caagatccgc cacaacatcg
     1261 aggatggcag cgtgcagctg gccgaccact accagcagaa tacccccatc ggcgatggcc
     1321 ctgtgctgct gcccgataac cactacctgt ccacccagag cgccctgtcc aaggacccca
     1381 acgagaagcg cgatcacatg atcctgctgg agttcgtgac cgccgccggc atcacccacg
     1441 gcatggacga gctgtacaag tgaggactag ataactgaac ttgtttattg cagcttataa
     1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgtg ggaagacaat
     1621 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1681 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2041 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2101 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2221 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2281 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2341 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2401 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2461 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2521 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2581 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2641 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2701 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2761 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2821 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2881 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2941 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3001 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3061 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3121 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3181 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3241 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3301 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3361 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3421 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3481 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3541 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3601 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3661 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3841 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4501 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     4621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5821 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     5881 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     5941 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     6001 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     6061 cgtttaatac gactcactat agggagagag agagaattac cctcactaaa gggaggagaa
     6121 gcatgaattc cccagtggaa agacgcgcag gcaaaacgca ccacgtgacg gagcgtgacc
     6181 gcgcgccgag cgcgcgccaa ggtcgggcag gaagagggcc tatttcccat gattccttca
     6241 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
     6301 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
     6361 gttttaaaat tatgttttaa
//
