LOCUS       pTwist_cDNA3.1_u        5392 bp    DNA     circular SYN 26-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5392)
  AUTHORS   siwong
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
COMMENT     Sequence Label: pTwist_cDNA3.1
FEATURES             Location/Qualifiers
     source          1..5392
                     /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"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    701^702
                     /label=Your gene here
     polyA_signal    752..976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1022..1450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1464..1794
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1645..1780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1861..2655
                     /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"
     polyA_signal    2831..2952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3001..3017)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3025..3041
                     /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(3049..3055,3056..3073,3074..3079))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3094..3115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3403..3991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4162..4953,4954..5022))
                     /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"
     promoter        complement(5023..5127)
                     /gene="bla"
                     /label=AmpR promoter
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 acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc agcggccgct cgagtctaga
      721 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
      781 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      841 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      901 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
      961 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     1021 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1081 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1141 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     1201 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1261 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1321 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1381 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1441 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1501 caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1561 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1621 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1681 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     1741 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     1801 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     1861 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1921 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1981 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2041 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2101 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2161 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2221 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2281 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2341 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     2401 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2461 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2521 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2581 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2641 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     2701 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     2761 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2821 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2881 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2941 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3001 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3061 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3841 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     5161 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     5221 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     5281 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     5341 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
