LOCUS       40924_37493        6127 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 6127)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6127)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        413..616
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             661..1377
                     /codon_start=1
                     /label=AcGFP1
                     /note="Aequorea coerulescens GFP"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILL
                     EFVTAAGITHGMDELYK"
     polyA_signal    1395..1516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1617..2045
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2059..2388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2455..3246
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3423..3556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3593..3609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3617..3633)
                     /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(3641..3671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3686..3707)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3995..4580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4754..5611)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5612..5716)
                     /label=AmpR promoter
     promoter        5982..6000
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6076..6094
                     /label=tet operator
                     /note="bacterial operator O1 for the tetR and tetA genes"
ORIGIN
        1 gagctcggta ccaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctaaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
      661 atggccagca agggcgagga gctgttcacc ggcgtggtgc ccatcctggt ggagctggac
      721 ggcgatgtga atggccacaa gttcagcgtg agcggcgagg gcgagggcga tgccacctac
      781 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc ctggcccacc
      841 ctggtgacca ccctgagcta cggcgtgcag tgcttctcac gctaccccga tcacatgaag
      901 cagcacgact tcttcaagag cgccatgcct gagggctaca tccaggagcg caccatcttc
      961 ttcgaggatg acggcaacta caagtcgcgc gccgaggtga agttcgaggg cgataccctg
     1021 gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg atggcaacat cctgggcaat
     1081 aagatggagt acaactacaa cgcccacaat gtgtacatca tgaccgacaa ggccaagaat
     1141 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg atggcagcgt gcagctggcc
     1201 gaccactacc agcagaatac ccccatcggc gatggccctg tgctgctgcc cgataaccac
     1261 tacctgtcca cccagagcgc cctgtccaag gaccccaacg agaagcgcga tcacatgatc
     1321 ctgctggagt tcgtgaccgc cgccggcatc acccacggca tggacgagct gtacaagtga
     1381 ggactagata actgaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1441 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1501 tcatcaatgt atcttatcat gtctgtggga agacaatagc aggcatgctg gggatgcggt
     1561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2041 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2401 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4501 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     4561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4861 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5161 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5401 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5701 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     5761 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5821 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     5881 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     5941 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt ttaatacgac tcactatagg
     6001 gagagagaga gaattaccct cactaaagat atttgcatgt cgctatgtgt tctgggaaat
     6061 caccataaac gtgaaactct atcattgata gagttcttat aagttctgta tgagaccact
     6121 cggatcc
//
